The effects of attenuated Salmonella carrying IL-2/NK4 double gene on immune function in mice / 中国生物制品学杂志

@#Objective To study the effects of attenuated Salmonella(Ty21a-pIRES-IL-2-NK4,TPIN)carrying interleukin-2(IL-2)/4-kringle antagonist of hepatocyte growth factor(NK4)double gene on humoral and cellular immune function.Methods Eighteen BALB/c mice,half male and half female,were randomly divided into control group(1. 5 mL 10%NaHCO3 by gastric tube feeding),Ty21a group(0. 1 mL Ty21a by gastric tube feeding)and TPIN group(0. 1 mL TPIN by gastric tube feeding),with 6 mice in each group. The immunization was boosted twice 7 d after the initial immunization. At 21d after administration,the blood samples were collected from eyeballs and the serum was separated,which was detected for the serum IgG antibody level by ELISA. The thymus and spleen of mice were isolated aseptically,and the spleen cells were stimulated by Ty21a and TPIN respectively in vitro. After 72 h,the proliferation ability of spleen cells was measured by CCK-8 assay,and the expression level of cytokines in spleen cells was detected by ELISA. The spleen and thymus were weighed,the spleen and thymus indexes were analyzed,and HE staining was performed.Results Compared with the control group and Ty21a group,the serum IgG level(F = 111. 74,P < 0. 01)and the contents of IFNγ,IL-4 and IL-10 in spleen cell supernatant(F = 38. 21,11. 37 and 26. 92,respectively,each P < 0. 05)increased significantly,as well as the spleen and thymus indexes(F = 10. 419 and 5. 859,respectively,each P < 0. 05)showed significant increase. In mice of Ty21a and TPIN group,the thymus cortex widened,lymphocytes increased,and there was mild inflammatory reaction;the white pulp and lymphocytes in spleen increased with neutrophil infiltration.Conclusion TPIN has a good immune protective effect,and can significantly stimulate the body to produce humoral immunity and cellular immunity,which may have a good therapeutic effect on tumors.

Immunogenicty and immunoprotective effect of Mycobacterium tuberculosis Ag85B-Fc2a DNA vaccine / 中国生物制品学杂志

@#Objective To evaluate the immunogenicity and protective effect of Mycobacterium tuberculosis(M.tb) Ag85B-Fc2a DNA vaccine in mice,so as to provide experimental basis for the development of the vaccine.Methods The recombinant plasmid pcD-Ag85B-Fc2a was identified by double digestion and sequencing,and then transfected into CHO-K1 cells.The expression of fusion protein Ag85B-Fc2a was detected by Western blot.Vector pcDNA3.1(+) and recombinant plasmid pcDAg85B-Fc2a were injected into female C57BL/6J mice through thigh muscle respectively(control group and immunization group),with 10 mice in each group,and booster immunization was carried out two weeks after the initial immunization.At14 d,28 d and 42 d after the initial immunization,serum samples was separated,and the titers of IgG antibody in the serum were detected by indirect ELISA.At 42 d after the initial immunization,the spleen of mice was taken aseptically and made into single cell suspension,and the proportions of CD4~+and CD8~+T cells were detected by flow cytometry.The remaining mice were injected with M.tb H37Ra through the tail vein 42 d after the initial immunization at a dose of 10~6 CFU/mouse.After 28 d of challenge,the lung and spleen of mice were collected aseptically.The number of bacteria in the left lung and spleen was measured by plate method,and the bacteria in the right lung was detected by auramine O fluorescence staining.Results Double digestion and sequencing results showed that the recombinant plasmid pcD-Ag85B-Fc2a was constructed correctly.After transfection into CHO-K1 cells,the fusion protein Ag85B-Fc2a with a relative molecular mass of about 70 000was detected.The Ag85B-specific IgG antibody titer in serum of mice in the immunization group was 1:3 200,1:12 160,and 1:12 800 at 14 d,28 d,and 42 d after the initial immunization,respectively,but no antibody titer was detected in the serum samples of control group.At 42 d after the initial immunization,the percentages of CD4~+ T cells in mouse spleen of control group and immunization group were(23.61±0.64)% and(26.92±0.80)%,and the percentages of CD8~+T cells were(14.12±0.87)% and(18.78±0.94)%,respectively,with significant differences(t=3.23 and 3.64,respectively,each P <0.05).After infection with M.tb H37Ra for 28 d,the numbers of bacteria were(4.73±0.13) and(3.81±0.14)CFU in the left lung,and(5.02±0.19) and(4.30±0.13) C.FU in the spleen of control group and immunization group,respectively,with significant differences(t=4.65 and 3.12,respectively,each P <0.01).The bacteria loading in the right lung was consistent with that in the left lung.Conclusion Ag85B-Fc2a DNA vaccine can induce specific humoral and cellular immune effects in mice,and can produce good protective effect against M.tb H37Ra infection.

Cellular and humoral immune status of thalassemia children with repeated blood transfusion in Yunnan province / 中国输血杂志

【Objective】 To investigate the changes in cellular immunity (peripheral blood lymphocyte subsets) and humoral immunity (serum immunoglobulin and ferritin) status among children with thalassemia who received repeated transfusions in Yunnan. 【Methods】 Forty-six children with thalassemia who underwent repeated blood transfusions from January 2020 to October 2022 were selected as the observation group. Forty children with thalassemia who did not receive blood transfusion were included in control group 1, and 46 healthy children underwent physical examination were included in control group 2. The differences in lymphocyte subsets, serum immunoglobulin levels and ferritin concentrations were compared among the three groups. 【Results】 For lymphocyte subsets: CD3+, CD4+ and CD4+/CD8+ in the observation group was lower than the control group 1 and 2: 57.60±8.36 vs 64.57±7.56 vs 66.58±5.65, 33.16±5.67 vs 38.62±8.36 vs 38.62±6.41 and 1.49±0.09 vs 2.32±0.15 vs 2.13±0.16, respectively; CD16+ CD56+ in the observation group was lower than the control group 2: 11.21±5.06 vs 16.70±7.92; CD8+ in the observation group was higher than control group 1 and control group 2: 26.63± 1.75 vs 20.60±1.43 vs 18.92±0.84; CD19+ in the observation group was higher than the control group 2: 24.06±6.42 vs 19.67 ±8.42, P<0.05, but no significant difference was noticed between the two control groups(P>0.05). For serum immunoglobulin and ferritin: IgG and ferritin in the observation group were higher than control group 1 and control group 2: 10.59±3.88 vs 7.02±3.88 vs 5.58±1.98 and 2 037.37±1 377.59 vs 72.63±56.71 vs 59.48±33.88. IgA in the observation group was higher than the control group 2: 1.06±0.92 vs 0.39±0.32(P<0.05), but no significant difference was noticed between the two control groups (P>0.05). The difference of IgM and IgE between the three groups was not significant (P > 0. 05). 【Conclusion】 The proportion of lymphocyte subsets in thalassemia children with repeated blood transfusion was imbalanced,and the level of immunoglobulin in humoral immunity was abnormal.

Orchestrating antigen delivery and presentation efficiency in lymph node by nanoparticle shape for immune response

Activating humoral and cellular immunity in lymph nodes (LNs) of nanoparticle-based vaccines is critical to controlling tumors. However, how the physical properties of nanovaccine carriers orchestrate antigen capture, lymphatic delivery, antigen presentation and immune response in LNs is largely unclear. Here, we manufactured gold nanoparticles (AuNPs) with the same size but different shapes (cages, rods, and stars), and loaded tumor antigen as nanovaccines to explore their disparate characters on above four areas. Results revealed that star-shaped AuNPs captured and retained more repetitive antigen epitopes. On lymphatic delivery, both rods and star-shaped nanovaccines mainly drain into the LN follicles region while cage-shaped showed stronger paracortex retention. A surprising finding is that the star-shaped nanovaccines elicited potent humoral immunity, which is mediated by CD4+ T helper cell and follicle B cell cooperation significantly preventing tumor growth in the prophylactic study. Interestingly, cage-shaped nanovaccines preferentially presented peptide-MHC I complexes to evoke robust CD8+ T cell immunity and showed the strongest therapeutic efficacy when combined with the PD-1 checkpoint inhibitor in established tumor study. These results highlight the importance of nanoparticle shape on antigen delivery and presentation for immune response in LNs, and our findings support the notion that different design strategies are required for prophylactic and therapeutic vaccines.

Effect of amantadine dimer adjuvant on humoral immune response induced by SARS-CoV-2 protein vaccine in mice / 中国生物制品学杂志

Effect of amantadine dimer adjuvant on humoral immune response induced by SARS-CoV-2 protein vaccine in mice@#Objective To investigate the effect of amantadine dimer adjuvant on humoral immune response induced by SARS-CoV-2 crown protein vaccine in mice.Methods The amantadine dimer was synthesized by substitution reaction ligation,hydrolytic acidification reaction ligation and amide condensation reaction ligation,with which as adjuvant,female BALB/c mice were immunized with the receptor-binding domain(RBD).The mice were randomly divided into five groups,six for each as follows:R6A+RBD group[21 μg(0.033 μmol)amantadine dimer+10 μg RBD],Ada+RBD group[10 μg(0.066 μmol)amantadine+10 μg RBD],Alu+RBD group(35 μg aluminum adjuvant+10 μg RBD),RBD group(10 μg RBD)and Blank group(0.9% normal saline),which were immunized i.m.on day 0,14 and 28 respectively.Serum samples were collected from tail vein of mice 7 d after the second dose and 14 d after the last dose and determined for specific IgG antibody levels by ELISA.Results The amantadine dimer was purified by thin layer chromatography(TLC)and identified by electrospray ionization-MS(ESI-MS)positive/negative ion mode.After two times of immunization,the antibody levels in sera at various dilutions of mice in R6A+RBD group were all higher than those of Ada+RBD group,while lower than those of Alu+RBD group.However,after three times of immunization,the antibody levels in sera at various dilutions of mice in R6A+RBD group were all significantly higher than those of Ada+RBD and Alu+RBD groups(each F > 30,each P < 0.000 1 and each P < 0.01).Conclusion Amantadine dimer adjuvant enhanced humoral immune response induced by SARS-CoV-2 protein vaccine in mice with good adjuvant effect,which may be used as an alternative adjuvant.This strategy based on existing drug transformation provided a new idea for the development of novel adjuvants.

Effect of endoscopic inflation-free thyroid surgery on patient-specific immune function and inflammatory response / 中国综合临床

Objective:To investigate the effect of inflation-free thyroid surgery on patient-specific immune function and inflammatory response.Methods:Sixty patients who underwent axillary endoscopic thyroid surgery at the First Affiliated Hospital of Bengbu Medical College from January 2021 to May 2023 were selected and randomly divided into an observation group and a control group using a random number table method, with 30 patients in each group. The control group underwent unilateral lobectomy and isthmus resection under transareola carbon dioxide inflation endoscopy, while the observation group underwent unilateral lobectomy and isthmus resection under transareola non inflation endoscopy. Compare the cellular immune related indicators, humoral immune related indicators, inflammatory response related indicators, as well as arterial blood partial pressure of carbon dioxide (PaCO 2) and end-expiratory carbon dioxide (PetCO 2) levels at T 1, T 2, and T 3 time points before anesthesia induction (T 1), during adenoidectomy (T 2), at the end of surgery (T 3), on the first postoperative day (T 4), and on the second postoperative day (T 5) in two groups of patients. The measurement data is represented by xˉ±s, and independent sample t-test is used for comparison between the two groups; The comparison between two groups at multiple time points was conducted using two factor analysis of variance, and the pairwise comparison was conducted using LSD- t test; Counting data is represented as an example (%), and inter group comparisons are made using χ 2 Inspection. Results:At time point T 1, there was no statistically significant difference between the two groups of patients in terms of cellular immune related indicators, humoral immune related indicators, and inflammatory response related indicators (all P>0.05). At time points T 2, T 3, T 4, and T 5, the CD3 +, CD4 +, CD4 +/CD8 + values and serum IgA, immunoglobulin A, immunoglobulin IgM The levels of immunoglobulin IgG were all lower than the T 1 time point in this group [control group: (31.49±5.37)%, (26.76±6.11)%, (34.75±5.99)%, (38.92±5.37)%, (51.78±5.90)%, (25.37±8.23)%, (19.12±7.13)%, (29.15±9.85)%, (33.49±8.03)%, (40.12±6.05)%, (0.97±0.28), (0.71±0.30), (1.11±0.36), (1.21±0.39)%, (1.69±0.41), (0.95±0.13), (0.91±0.14) (0.82±0.13), (0.96±0.16) g/L vs (1.21±0.20) g/L, (7.74±1.26), (7.33±1.31), (7.16±1.28), (7.82±1.31) g/L vs (9.18±1.52) g/L, (0.87±0.14), (0.86±0.13), (0.73±0.16), (0.88±0.15) g/L vs (1.16±0.22) g/L; Observation group: (35.82±5.71)%, (30.85±5.86)%, (39.43±5.68)%, (42.53±5.64)% vs (51.36±6.28)%, (30.39±9.76)%, (23.34±8.64)%, (34.68±11.37)%, (38.92±9.82)% vs (40.75±5.68)%, (1.15±0.35a), (0.89±0.38), (1.31±0.33), (1.52±0.37) vs (1.63±0.35), (1.04±0.17), (0.98±0.17) 0), (0.91±0.11a) (1.07±0.14) g/L vs (1.24±0.18) g/L, (8.51±1.35), (8.07±1.32), (7.93±1.34), (8.56±1.39) g/L vs (9.12±1.47) g/L, (0.95±0.11), (0.93±0.12), (0.83±0.18), (0.97±0.14) g/L vs (1.19±0.21) g/L], The CD8+values of both groups of patients were higher than those of the T 1 time point in this group, and at the T 4 time point, the control group was higher than the observation group [(29.89±8.99)% vs (25.70±6.91)%], with statistically significant differences (both P<0.05). At time points T 2, T 3, T 4, and T 5, both groups of patients had serum IL-interleukin-1 levels β、Interleukin IL-6, TNF tumor necrosis factor α The levels of CRP and CRPC reactive protein were higher than those at T 1 time point in this group [control group: (3.92±1.80), (4.16±1.86), (5.81±2.14), (4.46±1.87) ng/L vs (1.36±0.61) ng/L, (5.76±2.78), (6.68±3.12), (9.73±3.12), (4.65±2.78) ng/L vs (0.92±0.60) ng/L, (1.02±0.42), (1.30±0.61), (7.82±2.28), (6.65±2.16) mg/L vs (0.57±0.16) mg/L, (4.48±2.04) (4.48±2.04), (6.45±2.52), (5.33±2.15) ng/L vs (2.86±1.03) ng/L; Observation group: (3.04±1.09), (3.29±1.14), (4.56±2.01), (3.52±1.34) ng/L vs (1.65±0.63) ng/L, (4.12±2.11), (5.07±2.98), (8.07±3.15), (3.22±2.69) ng/L vs (0.98±0.53) ng/L, (0.81±0.34), (1.00±0.50), (6.65±2.03), (5.43±1.93) mg/L vs (0.56±0.12) mg/L, (3.39±1.81), (3.89±1.81) 4±1.93), (5.11±2.10) (3.96±2.03) ng/L vs (2.91±1.09) ng/L], and the control group was higher than the observation group, with statistically significant differences (all P<0.05). At time point T 1, there was no statistically significant difference in PaCO 2 and PetCO 2 between the two groups of patients (both P>0.05); At time points T 2 and T 3, the levels of PaCO 2 [(44.1±4.1), (45.8±4.0) mmHg] and PetCO 2 [(40.8±4.0), (42.1±3.5) mmHg] in the control group were higher than those at time points T 1 [(38.4±1.8), (36.3±1.9) mmHg] and observation group [PaCO 2: (38.3±2.0), (38.6±2.6) mmHg; PetCO 2: (36.3±1.9), (36.5±2.9) mmHg] (all P<0.05), There was no statistically significant difference between the observation group and this group at T 1 time point (all P>0.05). Conclusions:Inflation-free lumpectomy thyroid surgery is worthwhile as it has less suppressive effect on specific immunity and causes less inflammatory response compared to inflatable lumpectomy thyroid surgery.

Identificação, caracterização e determinação de padrões da resposta imune de epítopos imunogênicos de Plasmodium vivax / Identification, characterization and determination of immune response patterns of immunogenic epitopes of Plasmodium vivax

O Plasmodium vivax representa um grande desafio no controle da malária devido a sua vasta distribuição ao redor do globo, grande frequência de infecções sub microscópicas e habilidade de induzir recaídas em consequência das formas evolutivas que podem ficar latentes no fígado por longos períodos (hipnozoítos). O recente aumento de cepas de P. vivax resistentes aos fármacos disponíveis, a evolução de formas mais virulentas do parasito e a produção precoce de gametócitos, característica desta espécie, contribuem para classificar a malária vivax como um problema de saúde pública que merece atenção. Ainda que reconhecido por suas características biológicas peculiares e pelo agravamento recente de sua virulência, poucos investimentos têm sido feitos no desenvolvimento de ferramentas de controle para vivax. Portanto, o presente estudo teve como objetivo identificar e caracterizar novos alvos potenciais utilizando amostras de diferentes áreas endêmicas ao redor do mundo (Brasil, Mali, Camboja e Estados Unidos da América). Para tanto, investigamos e caracterizamos uma proteína recém descoberta na urina de pacientes naturalmente infectados (PvVir14); e descrevemos o potencial imunogênico de epítopos de células B de uma das proteínas mais bem caracterizadas de P. vivax, a PvAMA-1. Anti-IgG circulantes contra PvVir14 apareceram em 61% e 34.5% dos indivíduos do Brasil e Camboja, respectivamente, enquanto que indivíduos de Mali (infectados com falciparum e não expostos a vivax), tiveram 0% de reconhecimento. Ainda, os níveis de anti-PvVir14 correlacionaram-se com aqueles contra outros antígenos de vivax já bem caracterizados, como a PvCSP e a PvDBP, que foram reconhecidos por 7.6% e 42% dos indivíduos respectivamente. Com relação ao perfil celular, indivíduos sororreativos para PvVir14 apresentaram níveis significativamente maiores de células B atípicas circulantes (CD 21- CD 27-), sugerindo que tal tipo celular possa estar ligado à resposta anti-PvVir14. Entre as células T, os níveis de CD4+ e CD8+ diferiram entre indivíduos com e sem anticorpos contra PvVir14 (menor e maior expressão, respectivamente), enquanto que os níveis de células NKT foram mais expressivos em indivíduos sem anti-PvVir14. Se tratando da PvAMA-1, a antigenicidade dos peptídeos com epítopos para células B previamente selecionados foi avaliada através de múltiplos ensaios sorológicos utilizando amostras de indivíduos com infecção aguda por P.vivax do norte do Brasil. Os peptídeos sintéticos foram reconhecidos por 45.5%, 48.7% e 31.2% (PI, PII e PII, respectivamente) dos indivíduos selecionados para o estudo. Quando sintetizados em conjunto (tripeptídeo), a reatividade aumentou para 62%, porcentagem comparável àquela obtida pela proteína em sua forma e tamanho originais (57%). Além disso, a reatividade anti-IgG conta o tripeptídeo foi reduzida em 42% pós-depleção, indicando que tais epítopos podem ser responsáveis por parte considerável da imunogenicidade da proteína. Esses resultados representam uma excelente perspectiva na identificação de novos alvos com potencial imunogênico para compor uma vacina, ou auxiliar no desenvolvimento de outras medidas de controle, como testes diagnósticos, já que contemplar diversos alvos do ciclo de vida do parasito parece ser a chave para alcançar a resposta robusta e protetora que uma vacina contra a malária vivax precisa para ter sucesso.

Plasmodium vivax is a major challenge for malaria control due to its wide geographic distribution, high frequency of submicroscopic infections, and ability to induce relapses due to the latent forms present in the liver (hypnozoites). The recent increase in drug-resistant P. vivax strains, the evolution toward more virulent forms and the early production of gametocytes adds up to make P. vivax malaria a public health issue of increasing importance. Besides its tricky biological features and new awareness of its virulence, minimal investments have been made in vaccine discovery for P. vivax. Given that, this study aimed to discover and characterize potential new targets for future vaccine development using samples from different endemic areas around the world (Brazil, Mali, Cambodia and United States of America). For this purpose, we investigated and characterized a novel protein recently discovered in the urine of naturally infected subjects (PvVir14) and described the immunogenic potential of peptides from a well-known vivax protein (PvAMA-1), which has been proved to have important B cell epitopes that can induce specific immune response. Circulating antibodies against PvVir14 appeared in 61% and 34.5% of subjects from Brazil and Cambodia, respectively, versus none (0%) of the P. falciparum-infected subjects from Mali who have no exposure to P. vivax. PvVir14 antibody levels correlated with those against other well-characterized sporozoite/liver (PvCSP) and blood stage (PvDBP-RII) antigens, which were recognized by 7.6% and 42% of Brazilians, respectively. Concerning the cellular immune profiling of Brazilian subjects, PvVir14 seroreactive individuals displayed significantly higher levels of circulating atypical (CD21− CD27−) B cells, raising the possibility that atypical B cells may be contribute to the PvVir14 antibody response. Among T cells, CD4+ and CD8+ levels differed (lower and higher, respectively) between subjects with versus without antibodies to PvVir14, while NKT cell levels were higher in those without antibodies. As for PvAMA-1, the antigenicity of the selected B-cell peptides was assessed by multiple serological assays using sera from acute P.vivax infected subjects. The synthetic peptides were recognized by 45.5%, 48.7% and 32.2% of infected subjects for peptides I, II and III respectively. Moreover, when synthetized together (tripeptide), the reactivity increases up to 62%, which is comparable to the reactivity found against the whole protein PvAMA-1 (57%). Furthermore, IgG reactivity against the tripeptide after depletion was reduced by 42%, indicating that these epitopes may be responsible for a considerable part of the protein immunogenicity. These results represent an excellent perspective on discovering new targets with immunogenic potential to compose a vaccine, or even to assist the development of other control measures, such as diagnostic tools, since contemplating several targets seems to be the key to achieving a robust and protective response that a malaria vaccine needs to be successful.

Immunity Status of Health Care Workers Post-Recovery from COVID-19: Natural Adaptive Immunity Persists at Nine Months Post-Infection : An Online Longitudinal Panel Survey

Background : Various studies have pinned longevity of protective Immunoglobulin-G (IgG) titres at 2-5 months. The robustness and longevity of the IgG antibody response to COVID-19 infection has been gauged in a cohort of 214 single institutional health care workers by serial quantitative immunometric tests. Currently no separate guidelines exist for vaccination of COVID-survivors and this study provides data to fill this lacuna in knowledge. Methodology : Prospective longitudinal panel survey administered to the same cohort of Health Care Workers (HCW) till such time they got vaccinated under Indian Government’s free vaccination drive for HCW. Depending upon the date of contraction of infection the HCW could be longitudinally monitored for variable periods (2-9 months). The survey questionnaire comprising multiple-choice, dichotomous, matrix and Likert-scale questions was deployed to the respondents online via email/WhatsApp. Data was expressed as box-whisker plots, trendlines and trend areas. A p-value<0.05 was considered statistically significant. The composite index of ‘Effective Immunity’ was calculated. Results : The mean IgG antibody titre was 11.13±8.6AU at 1-2m, 9.68±8.9AU at 3-4m, 8.35±5.9 AU at 6-7m and 7.87±4.4 AU at 8-9m after first symptom, respectively. The lowest titre at all time points was 0 while the highest titres were 46.8 AU, 56.5 AU, 23.4 AU and 17.4 AU at 1-2m, 3-4m, 6-7m and 8-9m, respectively. Conclusion : Adaptive active immunity acquired through natural infection may last for at least 9 months post-initial exposure and lies in the moderate protection range in 77% HCW, which can be extrapolated to vaccination and immunity passports. Separate vaccination guidelines are required for COVID-survivors. The first shot of vaccine serves as a booster second exposure/booster dose in all COVID-survivors.HCW with low IgG-titre may suffer from a false sense of security. Periodic quantitative IgG-titre based serological tests can help guide timing of second shot of vaccination and predict likelihood of re-infection

The clinical characteristics of delayed antibody production in mycoplasma pneumoniae pneumonia in children / 中国小儿急救医学

Objective:To clarify the clinical characteristics and related fators of children with delayed antibody production of mycoplasma pneumoniae pneumonia(MPP).Methods:Two hundreds and eithty-five cases of children hospitalized at Children′s Hospital of Soochow University with MPP(positive for nucleic acid testing of respiratory secretion)were chosen from January 1st, 2019 to September 31st, 2019.Delayed antibody production group included 36 cases, who were tested for negative IgM antibody meanwhile the titer of IgG antibody changed less than 4 folds within 14 days.Positive group included 249 cases who were tested for positive IgM antibody or the titer of IgG antibody changed over 4 folds within 14 days.The characteristics of clinical manifestation, immunology and radiology were comparatively analyzed.Results:The medium age of delayed antibody production group was 0.75(0.30, 2.78)years old, which was obviously younger than that from positive group[5.50(3.73, 7.20)years old]( P<0.001). Low level of serum immunoglobulin IgG was the independent effect factor of delayed production for Mycoplasma pneumoniae antibody( P=0.037). When the serum immunoglobulin IgG level was lower than 7.155mmol/L, the sensitivity of predicting delayed production for mycoplasma pneumoniae antibody would be 0.819 and the specificity was 0.833.The underlying diseases associated with delayed antibody production were hospitalization history during neonatal period( P=0.007)and congenital heart disease( P=0.001). There were 11.11%(4/36)of children appearing spasmodic cough, 41.67%(15/36)of children showing wheezing and 33.33%(12/36)showing diarrhea in delayed antibody group, which were significantly higher than those in positive group[0.40%(1/249), 24.50%(61/249)and 9.64%(24/249), respectively, P<0.05]. The incidence of fever in delayed antibody group were 63.89%(23/36), which was lower than that in positive group[92.37%(230/249)]( P<0.001), meanwhile, the fever last time was 2.50(0, 4.75)days in delayed antibody group, which was shorter than that in positive group[ 7(5.00, 8.50)days]( P<0.001). In the delayed antibody group, there was 19.44%(7/36)of children sufferring from lobar pneumonia, and no extrapulmonary manifestations occurred, which were significantly lower than those in positive group[75.50%(188/249), 14.86%(37/249)]( P<0.05). Conclusion:Delayed antibody production in children with MPP is more common when serum immunoglobulin IgG level is lower than 7.155 mmol/L, especially in the presence of neonatal hospital history and congenital heart disease.The clinical manifestations of these children are mainly characterized by spasmodic cough and wheezing, with low probability of fever, lobular pneumonia and extrapulmonary manifestations.

Construction of a novel bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 and evaluation of its immunogenicity in mice / 中华微生物学和免疫学杂志

Objective:To construct a bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 and to evaluate its immunogenicity in mice.Methods:The coding sequences for spike 1 (S1) protein of SARS-CoV-2 Beta variant and hemagglutinin (HA) of influenza A virus Cambodia (H3N2) strain were codon-optimized and synthesized. The two coding genes were ligated by the self-cleaving 2A peptide using over-lapping PCR to construct S1-2A-HA fragment, which was inserted into pVRC vector to construct the bivalent DNA vaccine, named as pVRC-S1-2A-HA. Indirect immunofluorescence assay (IFA) and Western blot were performed to detect the expression of S1 and HA proteins. BALB/c mice were immunized with pVRC-S1-2A-HA by intramuscular injection and electroporation. The humoral immune responses induced in mice were detected by indirect ELISA, pseudovirus neutralization assay and hemagglutination inhibition assay. Cellular immune responses were detected by IFN-γ ELISPOT, intracellular cytokine staining (ICS) and cytometric bead array (CBA).Results:The bivalent DNA vaccine pVRC-S1-2A-HA could express S1 and HA proteins in vitro. Specific cellular immune responses against S1 protein and specific IgG antibody against HA protein were significantly induced in mice with single-dose immunization. The antigen-specific immunity was significantly enhanced after booster immunization. The geometric mean titer (GMT) of specific IgG antibody increased to 3 251 for S1 protein and 45 407 for HA protein after two-dose immunization. Moreover, the S1-specific T cells increased to 1 238 SFC/10 6 cells. ICS results indicated that the booster vaccination induced CD4 + T and CD8 + T cells to produce IL-2, IFN-γ and TNF-α in mice. The secretion of various cytokines including IL-2, IL- 4, IL-6, IL-10 and IFN-γ in mouse splenocytes was induced after single-dose immunization. Conclusions:A bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 was constructed and could induce S1- and HA-specific humoral and cellular immune responses in mice, suggesting the great potential of it for further development and application.

A trimeric RBD protein as a booster vaccine enhanced the humoral response and broad neutralization against SARS-CoV-2 in mice / 中华微生物学和免疫学杂志

Objective:To investigate the efficacy of a SARS-CoV-2 recombinant protein vaccine as a booster dose.Methods:A new immunogen, namely RBD-sc-trimer, was designed by tandem repeating of single receptor binding domain (RBD) of SARS-CoV-2 spike (S) protein to mimic the trimeric form of RBD presented by the virus. The RBD-sc-trimer protein was expressed as a His-tagged fusion protein using a baculovirus expression system and purified by nickel affinity column. The purified protein was identified by Western blot. Its in vitro binding activity to human angiotensin converting enzyme 2 (hACE2) was analyzed by ELISA. The immunogenicity of RBD-sc-trimer as well as RBD proteins of other forms including RBD dimer (RBD-Fc), RBD monomer (RBD) and S protein trimer (S trimer) as a booster dose was evaluated in BALB/c mice. Results:In terms of both binding and neutralizing antibodies against SARS-CoV-2, RBD-sc-trimer showed an immunogenicity that was superior to that of RBD-Fc and RBD and close to the level of S trimer. The antibody response induced by RBD-sc-trimer was characterized as Th1-biased. Moreover, it displayed a stronger cross-neutralization activity against SARS-CoV-2 Beta, Delta and Omicron variants. The titer of neutralizing antibody against Omicron induced by RBD-sc-trimer only decreased by 9.1 folds relative to the prototype strain, while the antibody response induced by RBD-Fc and S trimer decreased by 68.4 and 70.8 folds, respectively.Conclusions:The recombinant protein, RBD-sc-trimer, which was capable of eliciting stronger humoral response in mice as a booster dose and showed the superiority in raising cross-reactive antibodies against SARS-CoV-2 variants over non-trimeric RBD forms, should be considered as an optimal immunogen for the development of more effective SARS-CoV-2 vaccines.

Study on rAd5F35-SIVenvT vaccine in combination with rMVA-SIVenvT vaccine in mice / 中华微生物学和免疫学杂志

Objective:To analyze the immune response in mice after immunization with vaccine of rAd5F35-SIVenvT in combination with rMVA-SIVenvT to evaluate the efficacy of different immunization strategies.Methods:Two recombinant viruses were identified in vitro by PCR and Western blot. The BALB/c mice were immunized with homologous and heterologous immune strategies. The numbers of splenic lymphocytes secreting IFN-γ were measured by ELISPOT assay, meanwhile SIV gp120 antibody titer were measured by ELISA assay. Results:SIVenvT protein was expressed effectively by rAd5F35-SIVenvT and rMVA-SIVenvT in HEK293 cells. The specific immune response reached its peak at 4-week post first immunization, then decreased. SIV Env specific cellular immune response and SIV gp120 specific antibody could be detected at 4-16 weeks post first immunization. The specific cellular response was significant stronger in heterologous immunization group than homologous group at 4 week and 16 week. Furthermore, heterologous immunization induced significant higher titer of SIV gp120 antibody at 4 week than homologous group.Conclusions:Specific immune response induced by rAd5F35-SIVenvT in combination with rMVA-SIVenvT was stronger than homologous vector immunization. The results provided references for further study in nonhuman primates.

Humoral immune responses to different doses of bivalent norovirus vaccine in mice / 中华微生物学和免疫学杂志

Objective:To investigate the humoral immune response to GⅠ.1/GⅡ.4 norovirus (NoV) virus-like particle (VLP) vaccine of different doses in mice.Methods:The GⅠ.1/GⅡ.4 norovirus vaccine was diluted into four different concentrations, containing 50, 25, 8.3 and 2.8 μg/dose antigens, respectively. Aluminum hydroxide adjuvant was used in the control group. Ten 7-week-old BALB/c mice in each group were immunized intraperitoneally with 0.5 ml vaccine or adjuvant on 0 d and 21 d. Blood samples were collected on 35 d and the histoblood group antigen (HBGA) blocking titers of GⅠ.1 and GⅡ.4 antibodies in serum were detected. The differences in antibody levels between the groups were analyzed by SPSS23.0 software.Results:GⅠ.1 and GⅡ.4 HBGA blocking antibodies in 50, 25 and 8.3 μg/dose groups became positive on 35 d after the first dose vaccination. The geometric mean titers (GMT) of GⅠ.1 and GⅡ.4 HBGA blocking antibodies were 488 (95%CI: 249-955) and 489 (95%CI: 302-790) in 50 μg/dose group, 278 (95%CI: 106-728) and 738 (95%CI: 299-1 820) in 25 μg/dose group, 300 (95%CI: 158-570) and 486 (95%CI: 222-1 068) in 8.3 μg/dose group, respectively. The positive rates of GⅠ.1 and GⅡ.4 blocking antibodies in 2.8 μg/dose group were 40% and 70% and the GMT were 23 (95%CI: 10-51) and 85 (95%CI: 24-304), respectively. The GⅠ.1 and GⅡ.4 blocking antibodies in the control group were negative. Statistically significant differences in antibody levels were found between 50, 25, 8.3 μg/dose groups and the control group ( P<0.05), as well as in GⅠ.1 blocking antibodies between 50, 25, 8.3 μg/dose groups and 2.8 μg/dose group ( P<0.05). GⅡ.4 antibody level in 25 μg/dose group was statistically different from that in 2.8 μg/dose group ( P<0.05). Conclusions:GⅠ.1/GⅡ.4 norovirus VLP vaccine at (50-8.3) μg/dose could induce humoral immune response in mice.

Detection and significance of immune function in oral mucosa-associated diseases / 口腔疾病防治

@#The evaluation of immune function plays an important role in the diagnosis, treatment and prognosis of many diseases. To date, immune function detection includes cellular immunity, humoral immunity, and inflammatory markers. In this paper, the application of immune function detection in the diagnosis, differential diagnosis and treatment monitoring of various diseases was discussed; then, the application value of immune function detection in the diagnosis and treatment of three common oral mucosa-related diseases, including recurrent aphthous ulcer (RAU), oral lichen planus (OLP), and oral squamous cell carcinoma (OSCC), were reviewed combined with the literature and our research. Our research found that RAU patients present abnormal humoral immune function and obvious inflammatory reactions, whereas OLP and OSCC patients present mild inflammatory reactions and more serious abnormal cellular and humoral immune function, so the combined detection of immune function has a certain guiding value for the diagnosis and treatment of these diseases. Moreover, in the future, it is necessary to carry out a study on large sample, multicenter and multiindex joint detection to better clarify the role of immune dysfunction in the pathogenesis of various diseases and its mechanism, to establish the corresponding diagnostic model and prognostic prediction model, to find more effective treatment methods.

Effect of nabumetone on humoral immune responses in mice / Efeito da nabumetona nas respostas imunes humorais em camundongos

Nabumetone is used to reduce the pain and inflammation in rheumatoid arthritis. In the current study, immunomodulatory effect of Nabumetone is investigated in mice. The control group was administered normal saline orally as placebo. Nabumetone was administered orally via gavage in two treatment groups at 14mg/kg.b.w. doses and 28mg/kgb.w., respectively. Haemagglutination (HA) assay, Jerne hemolytic plaque and mice lethality assays were applied. In HA assay, the titer was significantly decreased in Nabumetone treatment groups (P< 0.001). In Jerne hemolytic plaque formation assay, there was a significant reduction (P< 0.001) in number of plaques in Nabumetone treated groups when compared with control. In mice lethality assay, there was a significant difference in mortality ratio of mice in control and Nabumetone treated groups (P< 0.001). Therefore, it is concluded that Nabumetone suppresses the humoral immune response in mice.(AU)

A nabumetona é usada na redução da dor e inflamação da artrite reumática. No presente estudo, o efeito imunomodulador é investigado em camundongos. O grupo de controle recebeu solução salina via oral como placebo. Nabumetona foi administrada oralmente via gavagem em dois grupos de tratamentos com doses de 14mg/kg.b.w. e 28mg/kgb.w., respectivamente. Foram realizados ensaios de hemaglutinação (HA), placa hemolítica de Jerne e letalidade dos camundongos. No ensaio HA, o grau foi significativamente menor nos grupos de tratamento com nabumetoma (P< 0.001). No ensaio de formação de placa hemolítica de Jerne houve redução significativa (P< 0.001) no número de placas em grupos tratados com nabumetoma comparado ao controle. No ensaio de letalidade dos camundongos houve diferença significativa no grau de mortalidade de camundongos no grupo de controle e grupos tratados com nabumetoma (P< 0.001). Portanto, conclui-se que a Nabumetoma suprime a resposta imune humoral em camundongos.(AU)

Study on immunological functions of heat shock protein 40 kDa of Schistosoma japonicum / 中国血吸虫病防治杂志

Objective To investigate the immunological functions of heat shock protein 40 kDa of Schistosoma japonicum (SjHSP40). Methods The homology of the SjHSP40 protein sequence was analyzed and the B and T cell epitopes of SjHSP40 were predicted using bioinformatics tools. The full-length SjHSP40 gene was amplified using a PCR assay, and cloned into the prokaryotic expression vector pGEX-6P-1, which was transformed into Escherichia coli BL-21. The protein expression was induced with isopropyl β-D-thiogalactoside (IPDG), and then, the recombinant protein was purified with glutathione-sepharose 4B resin to yield the fusion protein GST-SjHSP40, which was checked with SDS-PAGE and Western blotting. Following immunization with GST-SjHSP40, the serum levels of anti-SjHSP40 IgG antibody and IgG1 and IgG2a subtypes were detected in BALB/c mice using ELISA. In addition, the effect of SjHSP40 on CD4+ T-cell subset differentiation was examined using flow cytometry. Results SjHSP40 contained 7 potential B cell epitopes and multiple T cell epitopes (CTL epitopes and Th epitopes). The prokaryotic expression plasmid pGEX-6p-1-SjSHP40 was successfully constructed, and the fusion protein GST-SjHSP40 was obtained following IPDG induction and protein purification. Significantly higher serum levels of anti-SjHSP40 IgG, IgG1 and IgG2a antibodies were detected in mice immunized with GST-SjHSP40 than in other groups; however, SjHSP40 showed no remarkable effects on CD4+ T-cell subset differentiation. Conclusions SjHSP40 may induce specific humoral immune responses in mice; however, it does not affect the balance of Th immune responses. It is suggested that SjHSP40 may be a potential vaccine candidate.

Exposure to Leishmania spp. infection and Lutzomyia spp. in individuals living in an area endemic for visceral leishmaniasis in Brazil

Abstract INTRODUCTION: This study aimed to investigate human exposure to Leishmania spp. infection and sandflies in an area endemic for the disease. METHODS: The presence of antibodies specific for Leishmania spp. and saliva of Lutzomyia spp. and that of L. infantum DNA in blood were evaluated. RESULTS: Antibodies against Leishmania spp. and sandfly saliva were observed in 20.8% and 37.7% of individuals, respectively. DNA of Leishmania spp. was amplified from the blood of one patient. CONCLUSIONS: The results suggest that Leishmania spp. infection may be underdiagnosed in this area.

Algunos indicadores de la inmunidad celular y humoral en pacientes con linfangitis / Some indicators of cellular and humoral immunity in patients with lymphangitis

Introducción: La linfangitis es la inflamación de los vasos linfáticos producida por gérmenes patógenos, caracterizada por su recurrencia y el compromiso de su sistema inmune. Es frecuente en Cuba. Objetivo: Evaluar algunos indicadores de la inmunidad celular y humoral en pacientes con linfangitis. Métodos: Se realizó un estudio descriptivo, prospectivo y analítico en 75 pacientes divididos en tres grupos: sin linfangitis (referencia), con linfangitis en un primer episodio y con linfangitis recidivante; todos atendidos en el Instituto Nacional de Angiología y Cirugía Vascular. Las variables estudiadas fueron: edad, sexo, color de la piel y algunos parámetros de la inmunidad celular y humoral. Se utilizaron las pruebas no paramétricas chi cuadrado y t de Student para comparar los grupos entre sí. Resultados: Se observó un predominio de sexo femenino (n= 47, 62,7 por ciento); de edades superiores a los de 40 años (n= 61, 81,3 por ciento) y del color de piel blanca (n= 37, 49,3 por ciento). La obesidad, la insuficiencia venosa crónica y la Diabetes Mellitus fueron los factores de riego más frecuentes. El grupo con linfangitis recidivante, con respecto a los otros grupos, presentó alteraciones en la inmunidad humoral con concentraciones incrementadas (p = 0,007) de todas las inmunoglobulinas. No hubo variaciones significativas en la inmunidad celular. Conclusiones: Las alteraciones encontradas en la inmunidad celular y humoral de los pacientes con linfangitis, tanto en la primera crisis como en la recidiva, no son suficientes para sugerir que pudieran influir en los procesos sépticos asociados a esta afección(AU)

Introduction: Lymphangitis is the inflammation of the lymphatic vessels produced by pathogenic germs and characterized by its recurrence and the compromise of the immune system. It is frequent in Cuba. Objective: To evaluate some indicators of cellular and humoral immunity in patients with lymphangitis. Methods: A descriptive, prospective and analytical study was carried out in 75 patients divided into three groups: without lymphangitis (reference), with lymphangitis in a first episode, and with recurrent lymphangitis; all attended at the National Institute of Angiology and Vascular Surgery. The variables studied were: age, sex, color of the skin and some parameters of cellular and humoral immunity. The non-parametric chi square and Student's t tests were used to compare the groups among each other. Results: A predominance of females was observed (n= 47, 62.7 percent); ages over 40 years (n= 61, 81.3 percent) and white skin color (n= 37, 49.3 percent). Obesity, chronic venous insufficiency and diabetes mellitus were the most frequent risk factors. The group with recurrent lymphangitis, with respect to the other groups, presented alterations in humoral immunity with increased concentrations (p= 0.007) of all immunoglobulins. There were no significant variations in cellular immunity. Conclusions: The alterations in the cellular and humoral immunity of the patients with lymphangitis, both in the first crisis as in the recidive, are not enough to suggest that they may impact in the septic processes associated with this pathology(AU)

Lymphocytic Count in Premenopausal Women with Iron Deficiency Anemia

Introduction: Iron deficiency anemia (IDA) is the mostcommon nutritional deficiency worldwide. It can causereduced work capacity in adults and impact motor and mentaldevelopment in children and adolescents. Aim of the currentresearch was to study the lymphocytic count in premenopausalwomen with iron deficiency anemia.Material and methods: The study was conducted in theDepartment of General Pathology of the medical institution.For the study, we selected 100 pre-menopausal womenbetween the age group of 18-40 years who were diagnosedwith iron deficiency anemia and their hemoglobin level wasless than 10 g/dL. 100 pre-menopausal women with normalhemoglobin level were recruited after matching with thesubjects for control group. The patients with thalassemia,leukemia or any other chronic and autoimmune disease wereexcluded from the study. Laboratory evaluation of eachsubject was done.Results: The mean age of the patients in study group was 32.67years and in control group was 34.58 years. There were 100subjects in each group. Table 2 shows the mean lymphocytecount in peripheral venous blood in pre-menopausal womenwith Iron deficiency anemia and normal healthy women. Themean CD3+, CD4+, CD8+, and CD19+ lymphocyte countswere 1.66, 0.71, 0.66, 0.41, 1.18 X 109/L, respectively, instudy group, and 1.82, 0.59, 0.81, 0.31 and 1.59 X 109/L,respectively, for the control group.Conclusion: Within the limitations of the study, this can beconcluded that significant change in seen in the lymphocytecount in premenopausal women with iron deficiency anemia.

Analysis of immunity function of infants delivered by human immunodeficiency virus infected mothers / 中华实用儿科临床杂志

Objective To explore the physical development and immune function of infants without human immunodeficiency virus(HIV)infection who were delivered by HIV_infected mothers. Methods Two hundred and ninety_seven infants delivered HIV_infected mothers in Guangxi province from January 2008 to November 2011 were selected as observation group. According to whether infants had HIV infection or not,the children were further divided into the HIV_infection group and the infants in the non_HIV infection group according to the presence or absence of HIV infection,and the infants in the non_HIV infection group were divided into the antiretroviral drug(ART)treatment group and the non_ART treatment group according to whether the mother had used ART during pregnancy. Ninety_one healthy children born at the same time were selected as the healthy control group. The physical examination,T lympho_cyte subgroup analysis and humoral immunity test were performed on all infants. Results The weight and body length at birth of infants born from HIV_infected mothers were all significantly lower than those in the healthy control group [(2. 86 ± 0. 49)kg vs.(3. 15 ± 0. 52)kg;(47. 05 ± 2. 20)cm vs.(50. 01 ± 2. 58)cm],and the differences were sta_tistically significant(t﹦2. 652,2. 247,all P〈0. 05). The CD8 level and CD4∕CD8 ratio of infants delivered by HIV_infected mothers had no significant differences statistically compared with those in the healthy control group[(21. 31 ± 6. 49)% vs.(22. 01 ± 5. 43)%;1. 82 ± 0. 79 vs. 1. 82 ± 0. 67,t﹦0. 933,0. 033,all P〉0. 05];the CD3 and CD4 levels were lower than those in the healthy control group[(62. 36 ± 7. 94)% vs.(65. 70 ± 6. 32)%;(4. 83 ± 7. 62)% vs.(37. 02 ± 5. 69)%],and the differences were statistically significant(t﹦3. 66,2. 946,all P〈0. 01). The immunoglobulin(Ig)M,IgG and IgA levels of children born to HIV_infected mothers had no statistically significant differences compared with those in the healthy control group[(1. 79 ± 0. 66)g∕L vs.(1. 76 ± 0. 66)g∕L;(8. 96 ± 2. 74)g∕L vs.(8. 80 ± 1. 97)g∕L;(0. 85 ± 0. 57)g∕L vs.(0. 86 ± 0. 41)g∕L,t﹦0. 341,0. 619,0. 173,all P〉0. 05). The weight and body length at birth of non_HIV infected children born from HIV_infected mothers were all significantly lower than those in healthy control group[(2. 92 ± 0. 43)kg vs.(3. 15 ± 0. 52)kg;(49. 03 ± 2. 22)cm vs.(50. 01 ± 2. 58)cm],and the differences were statistically significant( F﹦4. 163,2. 87,all P〈0. 05). The birth weight,birth length and head circumference of the ART group were all significant lower than those in the healthy control group[(2. 90 ± 0. 43)kg vs.(3. 15 ± 0. 52)kg;(48. 27 ± 1. 89)cm vs.(50. 01 ± 2. 58)cm;(31. 80 ± 1. 47)cm vs. (34. 88 ± 3. 21)cm],and the differences were statistically significant( F﹦3. 711,2. 970,3. 689,all P〈0. 05). The CD8 level and CD4∕CD8 ratio of non _ HIV infected children born to HIV _ infected mothers had no significant differences statistically compared with those in the healthy control group[(20. 77 ± 5. 60)% vs.(22. 01 ± 5. 43)%, 1. 85 ± 0. 76 vs. 1. 82 ± 0. 67,F﹦43. 568,11. 705,all P〉0. 05];the CD3 and CD4 levels were lower than those in the healthy control group[(62. 27 ± 7. 94)% vs.(65. 70 ± 6. 32)%;(35. 30 ± 6. 86)% vs.(37. 02 ± 5. 69)%],and the differences were statistically significant(F﹦7. 083,28. 06,all P〈0. 05). Conclusions The humoral immune func_tion of the non_HIV infected infants delivered by HIV_infected mothers is not significantly affected,but the physical development at birth and cellular immune function are significantly affected. ART during pregnancy is not a major factor in the limitation of physical development at birth. Therefore,the nutrition support for the infants delivered by HIV_in_fected mothers and prevention of infection are especially necessary clinically.