ABSTRACT
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.
ABSTRACT
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.
Subject(s)
Plasmodium vivax , Immunity, Humoral , Malaria , Academic Dissertation , EpitopesABSTRACT
【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.
ABSTRACT
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
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
@#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.
ABSTRACT
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.
ABSTRACT
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)
Subject(s)
Animals , Mice , Immunity, Humoral/drug effects , Nabumetone/administration & dosage , Immunologic Factors/analysis , Arthritis, Rheumatoid/veterinary , Saline Solution , HemagglutinationABSTRACT
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.
Subject(s)
Humans , Animals , Male , Female , Adolescent , Adult , Aged , Young Adult , Psychodidae/parasitology , Insect Vectors/parasitology , Leishmania/isolation & purification , Leishmaniasis, Visceral/transmission , Brazil , Enzyme-Linked Immunosorbent Assay , Polymerase Chain Reaction , Leishmania/genetics , Leishmania/immunology , Leishmaniasis, Visceral/epidemiology , Middle AgedABSTRACT
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.
ABSTRACT
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)
Subject(s)
Humans , Male , Female , Risk Factors , Indicators and Reagents , Lymphangitis , Immunoglobulins , Epidemiology, Descriptive , Retrospective Studies , InflammationABSTRACT
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.
ABSTRACT
Objective: To explore the regulatory effect of Zilongjin Tablet on immune function of postoperative patients with lung cancer from humoral immunity and cellular immunity, and to seek an effective method for comprehensive treatment of lung cancer. Methods: Sixty patients with lung cancer after operation were divided into two groups according to the digital random table, 30 in the control group and 30 in the treatment group. The control group was given routine supportive treatment after operation. The treatment group was given Zilongjin Tablets (4 tablets, 3 times/d) with routine supportive treatment on day 3 after operation. The indexes of immunoglobulin (IgA, IgG, IgM), T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+), and NK cells were compared between the two groups. Results: On day 2 after operation, IgA, IgG, IgM, CD3+, CD4+, CD4+/CD8+ in the two groups decreased compared with those before operation, and the difference was statistically significant (P 0.05). On day 14 after operation, the levels of IgA, IgG, IgM, CD3+, CD4+, CD4+/CD8+, NK in the two groups increased, the NK of the control group and the IgA, IgG, IgM, CD3+, CD4+, CD4+/CD8+ of the treatment group were significant differences compared with the 2nd day after operation (P < 0.05). Moreover, there were significant differences in IgA, IgG, CD4+, CD4+/CD8+, NK between the two groups on day 14 after operation (P < 0.05). Conclusion: Patients with lung cancer have immunosuppressive status in the early postoperative period. Zilongjin Tablets can improve the postoperative humoral immunity and cellular immunity of patients with lung cancer, promote the recovery of immune function and enhance their anti-tumor ability.
ABSTRACT
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.
ABSTRACT
Theileria equi is an infectious hemoprotozoan agent of equine piroplasmosis, a disease that has severe economic and sanitary impact internationally. In addition to its common clinical features, piroplasmosis can cause gestational losses and neonatal damage, which makes neonates susceptible to this disease. The aim of this study was to evaluate the dynamics of humoral immune response to recombinant EMA-2 of T. equi in pregnant mares and foals, as well as the transfer of vaccine antibodies through the colostrum ingested by sucking foals. For vaccine production, the EMA-2 expression gene was cloned and expressed in the yeast species, Pichia pastoris. Thirty-six horses were used, of which 18 were pregnant mares and 18 were foals. The mares were divided into control and vaccinated groups, and the vaccinated group received three doses of rEMA-2 every 21 days starting at 300 days of gestation. Foals from vaccinated and control groups were evaluated until the sixth month of life. The production of antibodies by foals on the rEMA-2 vaccination schedule was also evaluated from the second month of life. Foals in the vaccinated group had received three doses of the vaccine every 21 days. The method used to evaluate serum and colostrum samples was indirect ELISA, and plates were sensitized with the rEMA-2 protein. At the end of the vaccination schedule, vaccinated mares showed a 2.3-fold increase in antibody levels when compared to baseline values. The colostrum of vaccinated mares presented antibody levels of 1.0432±0.33. Foals delivered by vaccinated mares presented levels of antibodies greater than those of foals delivered by control mares after their first time sucking (at about twelve hours after birth). Foals vaccinated in the second month of life showed an 8.3-fold increase in antibody levels when compared to baseline values. The vaccination schedule with rEMA-2 was able to stimulate humoral immunity in pregnant mares. Vaccine immunoglobins were concentrated in the colostrum of vaccinated mares and foals delivered by these mares showed an increase in serum levels of vaccine antibodies after the first-time sucking.(AU)
Theileria equi é um hemoprotozoário, agente da piroplasmose equina, doença de impacto sanitário e econômico internacional. Em éguas gestantes além da doença clínica, podem ocorrer abortos e danos ao neonato, caracterizando grande susceptibilidade à doença no período neonatal. O objetivo deste estudo foi avaliar a dinâmica da resposta imune humoral à EMA-2 recombinante de T. equi em éguas gestantes e potros, bem como a transferência de anticorpos vacinais no colostro. Foram utilizados 36 equinos, sendo 18 éguas gestantes e 18 potros. As éguas foram divididas em grupo controle e vacinado, que receberam rEMA-2 a partir dos 300 dias de gestação em três doses com intervalos de 21 dias. Para produção da vacina, o gene de expressão de EMA-2 foi clonado e a proteína expressa em Pichia pastoris. Os potros provenientes de éguas dos grupos vacinado e controle foram avaliados até o 6º mês de vida. Avaliou-se também a produção de anticorpos em potros submetidos ao esquema vacinal com rEMA-2 a partir do 2º mês de vida, que receberam três doses da vacina em intervalos de 21 dias. O método escolhido para a avaliação das amostras de soro e colostro foi ELISA indireto, com sensibilização pela proteína rEMA-2. Nas éguas gestantes vacinadas com rEMA-2 ocorreu o incremento de 2,3 vezes o valor basal ao final do esquema vacinal. O colostro de éguas vacinadas apresentou título médio de anticorpos de 1,0432±0,33, e potros provenientes de éguas vacinadas apresentaram média maior que os provenientes de éguas controle após a primeira mamada (12 horas). Os potros que passaram por esquema vacinal a partir do 2º mês de vida obtiveram incremento de 8,3 vezes o valor basal de anticorpos. O esquema vacinal com rEMA-2 foi capaz de estimular a imunidade humoral em éguas gestantes. Éguas gestantes vacinadas concentraram imunoglobulinas vacinais no colostro, e os potros provenientes destas obtiveram incremento nos níveis séricos de anticorpos vacinais após a primeira mamada.(AU)
Subject(s)
Animals , Female , Pregnancy , Vaccines/immunology , Horses/immunology , Horses/microbiology , TheileriaABSTRACT
Este trabalho teve como objetivo avaliar o perfil imunogênico em ovinos de três vacinas produzidas com linhagens brasileiras de Mycoplasma agalactiae. O perfil proteico do antígeno vacinal foi avaliado por SDS-PAGE e a imunogenicidade da vacina pela técnica de Western blot. A vacina foi inativada com formol, adsorvida em hidróxido de alumínio (Vacina 1), Montanide IMS-2215 (Vacina 2), Montanide Gel-01 (Vacina 3) e administradas em três doses. Entre a primeira e a segunda dose houve um intervalo de 21 dias, e entre a segunda e a terceira de 180 dias. O pool de soros de dez ovinos coletados nos períodos 0, 21, 35, 90, 150, 210, 270 e 360 dias pós-vacinação foram testados pela técnica de Western blot. A vacina 2 foi mais antigênica, com detecção de anticorpos 21 dias após a primeira dose. Para as vacinas 1 e 3, os anticorpos são verificados após 35 dias, com queda acentuada aos 90 dias; apenas anticorpos contra a proteína de 48 kDa apareceram após a terceira dose de forma discreta. Contra a vacina 2, ainda persistiram anticorpos contra as proteínas de 48, 55 e 80 kDa nos períodos 90, 150 e 210, que aumentaram após a terceira dose. Conclui-se que a vacina 2 induziu a resposta humoral de forma estável contra proteínas de M. agalactiae.
This study aimed to evaluate the immunogenic profile in sheep of three vaccines produced with a field Brazilian strains of Mycoplasma agalactiae. The vaccine protein profile was evaluated by SDS-PAGE and vaccine immunogenicity by Western blot. The vaccine was inactivated with formaldehyde and adsorbed onto three different adjuvants: with aluminum hydroxide (Vaccine 1), Montanide IMS 2215 (Vaccine 2), and Gel Montanide-01 (Vaccine 3). The vaccine was administered in three doses. Between the first and second dose there was an interval of 21 days, and between the second and the third one of 180 days. A pool of ten sera collected in 0, 21, 35, 90, 150, 210, 270 and 360 days after first vaccination were tested by Western blot techniques. The second vaccine was more antigenic with antibody detection 21 days after first dose. For both vaccines 1 and 3, antibodies were present 35 days after first dose, with a significant drop at 90 days; only antibodies against 48 kDa protein discreetly appeared after the third dose. Stimulation induced by vaccine 2 produced antibodies against 48, 55 and 80 kDa proteins that persisted until 90, 150 and 210 days after first dose, which rose again after third dose. It was concluded that the vaccine 2 induced stable humoral immunity against M. agalactiae proteins.
Subject(s)
Animals , Sheep , Vaccines , ImmunityABSTRACT
ABSTRACT Introduction: Immune response to vaccination in infants born prematurely may be lower than in infants born at full-term. Some clinical factors might be associated with humoral immune response. Objectives: The objectives of this study were to compare the immune response to measles and varicella vaccination in infants born prematurely with those born at full-term and to analyze factors associated with measles and varicella antibody levels. Methods: Prospective study including two groups of infants aged 12 months. One group of infants born prematurely with birth-weight <1500 g and who were in follow-up at the outpatient clinic for preterm infants at the institution and other group of infants born at full-term. Infants with malformations, primary immunodeficiency diseases, born to HIV-positive mothers or who had received plasma or immunoglobulin transfusions five months before or three weeks after vaccination were excluded. Plasma antibodies were measured by ELISA and factors associated with antibody levels were assessed by linear regression. Results: Sixty-five premature and 56 full-term infants were included. The percentage of immune individuals after vaccination against measles (100% vs. 100%) and varicella (92.5% vs. 93.2%) were similar in both groups, as well as the antibody levels against measles (2.393 vs. 2.412 UI/mL; p = 0.970) and varicella (0.551 vs. 0.399 UI/mL; p = 0.114). Use of antenatal corticosteroids decreased measles antibody levels whereas breastfeeding for more than six months increased varicella antibody levels. Conclusions: Humoral responses to measles and varicella were similar between infants born prematurely and full-term infants. Measles antibody levels were negatively associated with antenatal corticosteroid use; varicella antibodies were positively associated with prolonged breastfeeding.