Nefropatía por enfermedad relacionada a IGG4 / Nephropathy due to IGG4-related disease

IgG4-related disease (ER-IgG4) is a group of systemic fibro-inflammatory diseases, whose renal involvement is rare and difficult to diagnose. Diagnosis is usually made by serological and histological studies. Treatment is based on systemic corticosteroids. The renal prognosis is determined by the patient's comorbidities and the degree of fibrosis in the renal biopsy. We present the case of an elderly patient with exacerbated chronic kidney disease, whose study showed nephropathy associated with ER-IgG4.

La enfermedad relacionada a IgG4 (ER-IgG4) es un grupo de enfermedades fibro-inflamatorias sistémicas, cuya afectación renal es poco frecuente y de difícil diagnóstico. Habitualmente el diagnóstico se realiza mediante estudios serológicos e histológicos. El tratamiento se basa en corticoides sistémicos. El pronóstico renal está determinado por las comorbilidades del paciente y el grado de fibrosis en la biopsia renal. Se presenta el caso de un paciente adulto mayor con enfermedad renal crónica reagudizada, cuyo estudio demostró nefropatía asociada a ER-IgG4.

Expression of Gn protein of severe fever with thrombocytopenia syndrome virus and its immunogenicity / 中国生物制品学杂志

@#Objective To express the Gn protein of severe fever with thrombocytopenia syndrome virus(SFTSV) through adeno-associated virus 9(AAV9) expression system and evaluate its immunogenicity.Methods SFTSV Gn gene was inserted into viral vector pAAV-CMV-FH and the recombinant plasmid was transfected into HEK293T cells to obtain recombinant virus AAV9-Gn.The expression of Gn protein was determined by immunofluorescence and Western blot.Eighteen fernale BALB/c mice were randomly divided into three groups:Mock group(serum-free DMEM),AAV9-GFP group(1 × 10~(11) vg) and AAV9-Gn group(1 × 10~(11) vg),all of which were injected intramuscularly into the right hind limb at a dose of 100 μL per mouse.The body mass,diet,behavior and mental state of mice in each group were monitored continuously for 21 d,and the change rate of body mass was calculated;At 2,4,8 and 16 weeks after immunization,the levels of SFTSV neutralizing antibody in serum of mice in each group were detected by fluorescent reduction neutralization test(FRNT),and the levels of specific IgGl and IgG2a in serum of mice in AAV9-Gn group were detected by ELISA.Results After incubation with specific antibody,Vero cells transfected with AAV9-Gn showed specific green fluorescence under fluorescence microscope,and had specific binding to mouse anti-SFTSV Gn monoclonal antibody,and the specific binding bands were found at a relative molecular mass of about 61 000.The body mass of the three groups showed an increasing trend,there was no significant difference between the three groups(F=0.158—2.621,P> 0.05),and the diet,behavior and mental state were normal.At 2,4,8 and 16 weeks after immunization,the titer of SFTSV neutralizing antibody in serum of mice in AAV9-Gn group was significantly higher than that of Mock group and AAV9-GFP group(H=13.332—14.538,each P <0.001),and the titer peak appeared at 8 weeks;The level of specific IgG1 in serum of mice was significantly higher than that of IgG2a(F=4.373—12.975,each P <0.05) at different time points.Conclusion SFTSV Gn protein can be expressed correctly through AAV9 expression system,and has low toxicity to mice with good immunogenicity,which is expected to be a candidate component of SFTSV vaccine.

Preparation of anti-PD-L1 nanobodies fused with C3Fab and their effect on plasma half-life / 国际生物医学工程杂志

Objective:To prepare the anti-programmed death-ligand 1 (PD-L1) nanoantibody P3C8-C3Fab by ligating with C3Fab and to investigate its role in plasma half-life.Methods:The C3Fab peptide derived from protein G was molecularly fused with the nanobody P3C8 by DNA recombination technology. The nanoantibody P3C8-C3Fab was inducibly expressed and purified in the E. coli BL21 strain, and the binding of it to PD-L1 protein, mouse IgG, and PD-L1-expressing tumor cells was detected by enzyme-linked immunosorbent assay (ELISA). The residual P3C8-C3Fab was detected in mouse serum at different times using double-antibody sandwich ELISA to assess the prolongation of the plasma half-life of PD-L1 nanobodies by C3Fab. Results:The nanoantibody P3C8-C3Fab was successfully constructed, and it could efficiently express itself in soluble form in BL21. The purified NbP3C8-C3Fab protein was obtained with a mass fraction of about 90% at a yield of 7.18 mg/L. The affinity of P3C8-C3Fab for PD-L1 protein and mouse IgG gradually increased with increasing mass concentration and showed a concentration correlation. The binding of P3C8-C3Fab to lung cancer A549 cells showed a concentration correlation. The concentration standard curve of P3C8-C3Fab in mouse serum showed a typical S-shape with a concentration correlation. The plasma half-life of P3C8 was only 0.44 h, while the plasma half-life of P3C8-C3Fab was 21.27-fold higher, up to 9.36 h.Conclusions:The linkage of C3Fab to the nanobodies of P3C8 can significantly prolong the plasma half-life of P3C8, which is valuable for the improvement of in vivo nanobody effects.

Analysis of detection results of food-specific IgE and IgG4 in children with allergic diseases / 临床检验杂志

Objective To investigate the clinical significance of the detection of food-specific IgE(fsIgE)and IgG4(fsIgG4)antibod-ies in the diagnosis and treatment of children's allergic diseases.Methods A total of 641 children with allergic diseases visited Hang-zhou Women's Hospital during November 2022 and November 2023 and 100 healthy children during the same period were selected,and the levels of fsIgE and fsIgG4 in their serum samples were detected.Results The total positive rates of fsIgE and fsIgG4 in children with allergic diseases were 37.44%and 90.48%,respectively,which were significantly higher than the 11.00%and 61.00%of healthy children(P＜0.05).The main food allergens in children were milk and eggs.The intensity of fsIgE was concentrated in the 1-2 level and the positive result for a single food was more common.The intensity of fsIgG4 was concentrated in Grade 3 and the positive results for multiple foods were more common.The comparison of the detection results of sIgE and sIgG4 to the same food showed that the posi-tive rate of sIgG4 was significantly higher than that of sIgE(P＜0.05),and the consistency between them was poor(κ＜0.4).There were no significant differences in the positive rates of fsIgE and fsIgG4 between different genders(P＞0.05).The highest positive rate of fsIgE was seen in the infant group and the highest positive rate of fsIgG4 was seen in the childhood group(P＜0.05).The positive rates of sIgE to egg protein,shrimp,crab,and cashew were mainly seen in children with skin diseases(P＜0.05).The positive rates of sIgG4 to egg and milk were mainly seen in children with respiratory diseases(P＜0.05).The positive rates of sIgG4 to shrimp and wheat were mainly seen in children with skin diseases(P＜0.05).Analysis of allergen components showed that the β-lactoglobulin,α-whey protein,and casein in milk and mucin in eggs were important allergenic components.Conclusion Food allergy is an important factor leading to children's allergic diseases.The combined detection of fsIgE and fsIgG4 can provide experimental basis for clinical di-agnosis and treatment as well as personalized dietary guidance of children.

Clinicopathologic characteristics of Epstein-Barr virus-positive inflammatory follicular dendritic cell sarcoma:report of nine cases / 临床与实验病理学杂志

Purpose To explore the clinicopathologic fea-tures of Epstein-Barr virus-positive inflammatory follicular den-dritic cell sarcoma(EBV+IFDCS).Methods The clinico-pathologic features of 9 cases of EBV+IFDCS were retrospective-ly analyzed and followed up.Results The age of 9 patients with EBV+IFDCS ranged from 22 to 78 years(mean 44.7 years).7 cases occurred in the liver and 2 in the spleen.Fi-brinoid degeneration and hyaline degeneration in the vessel walls(6/9),eosinophilic infiltration(3/9),and epithelioid granulo-mas(2/9)were seen in some cases.The tumor cells expressed CD21(7/9),CD23(8/9)and CD35(9/9),partially ex-pressed SMA(6/9)and D2-40(1/9).It was noteworthy that 2 cases from the spleen accompanied by high expression of IgG4 plasma cells(80-135/10 HPF),and in the liver(0-36/10 HPF).All cases were followed up for 3-84 months,with 6 pa-tients disease-free,2 patients underwent metastasis,1 patient lost of follow-up.Conclusion EBV+IFDCS is a rare low-grade malignant tumor.EBER in situ hybridization and immunohisto-chemical detection play important roles in the diagnosis and dif-ferential diagnosis of EBV+IFDCS.Surgical resection is the main therapeutic intervention for EBV+IFDCS,and patients re-quire long-term post-surgical follow-up.

Clinical features and lymphocyte subtypes in patients with IgG 4-related diseases / 中华内科杂志

Objective:To deepen understanding of IgG 4-related diseases (RDs), we analyzed the associated lymphocyte subtypes, and explored the pathogenesis and potential immunotherapeutic targets. Methods:Eighty-six patients with IgG 4-RDs were enrolled, and their clinical characteristics, peripheral lymphocyte subtypes, and disease course were analyzed. Results:The mean age of the participants was 36-87(62±11) years; 51 were male (59.3%) and 35 were women (40.7%); and 34.9% had a history of allergy. Follow-up lasted 4.8 (0.4, 14.1) months. The most common symptoms were abdominal pain, and submandibular gland and lacrimal gland swelling (each 20.9%). Sixty-five (75.6%) participants had multiple organ involvement, and the most frequently affected organs were the pancreas (52.3%), submandibular gland (51.2%), and lacrimal gland (34.9%). A high eosinophil count; high IgE, IgG, IgG 1, and IgG 4 concentrations; and low complement C3 and C4 concentrations were present in 18.8% (16/85), 30.0% (24/80), 72.9% (62/85), 58.3% (28/48), 89.5% (77/86), 61.2% (52/85), and 50.0% (42/84), respectively, of the participants. In addition, 64.7% (55/85) were positive for autoantibodies, and the most frequent was anti-nuclear antibody (63.5%). The proportion of CD4 +T lymphocytes increased in 25.7% (9/35) of the participants, which was accompanied by an increase in the ratio of CD4 +/CD8 +T lymphocytes (22.9%, 8/35). Importantly, most participants (90.0%, 18/20) had a high proportion of regulatory T (Treg) cells. High interleukin (IL)-2, IL-6, and IL-10 concentrations were present in 50.0% (11/22), 33.3% (10/30), and 16.7% (5/30), respectively, of the participants. Substantial lymphoplasmacytic infiltration, fibrosis, IgG 4-positive plasma cell infiltration, and lymphoid follicle hyperplasia or ectopic formation were present in 79.2% (42/53), 67.9%(36/53), 35.8%(19/53) and 30.2% (16/53), respectively, of the participants. Fifty-three participants with detailed pathologic data were also further evaluated, of whom 24.5% (13/53), 3.8% (2/53), and 67.9% (36/53) had definite, probable, and possible diagnoses; and 3.8% (2/53) could not be diagnosed. Compared with baseline, the percentage of eosinophils and the IgE, IgG, and IgG 4 concentrations decreased significantly; and the complement C3 and C4 concentrations had increased significantly after 6 months of treatment (all P<0.05). The IgG 4 concentration after 6 months of treatment negatively correlated with that of C4, and positively correlated with the baseline concentration of IgE and the IgG 4/IgG ratio. Conclusion:IgG 4-RDs are a group of diseases characterized by male predisposition; multiple organ involvement; a high eosinophil count; high IgE, IgG, IgG 1, and IgG 4 concentrations; and a low C3 concentration. Peripheral CD4 +T cells and Treg cells are also more abundant. The diseases can be controlled with glucocorticoids and immunosuppressive drugs in the majority of instances. The IgG 4 concentration after 6 months of treatment negatively correlates with the baseline complement C4 concentration and positively correlates with the IgE concentration and IgG 4/IgG ratio, which suggests that IgG 4/IgG, IgE, and complement should be closely monitored to evaluate disease activity and the efficacy of treatment in such patients.

Clinical and endoscopic ultrasound features of IgG4-related autoimmune pancreatitis / 中华消化内镜杂志

Objective:To analyze the differences in clinical and endoscopic ultrasonography (EUS) findings between diffuse and focal IgG4-related autoimmune pancreatitis (IgG4-AIP).Methods:Data of patients diagnosed as having IgG4-AIP who underwent EUS at Chinese PLA General Hospital from September 2011 to April 2022 were retrospectively collected. General clinical data, EUS features, and postoperative pathology were analyzed for characteristic differences.Results:A total of 40 patients were included in the study, 60.03±10.87 years old, a higher proportion of males (85.0%, 34/40). All patients underwent EUS, and 28 underwent EUS-guided fine-needle aspiration. Among the 40 patients, 29 (72.5%) had diffuse type and 11 (27.5%) had focal type. Abdominal pain [65.5% (19/29) VS 18.2% (2/11), χ2=5.393, P=0.020] and thickening of the bile duct wall [51.7% (15/29) VS 9.1% (1/11), χ2=4.394, P=0.036] were more common in the diffuse type, while main pancreatic duct dilation [45.5% (5/11) VS 10.3% (3/29), χ2=4.146, P=0.042] was more common in the focal type, with the lesion most commonly located in the pancreatic head (90.9%, 10/11). There was no significant difference in the presence of chronic pancreatitis parenchymal changes between the two groups [34.5% (10/29) VS 27.3% (3/11), χ2=0.003, P=0.955]. Conclusion:There are certain differences in abdominal pain and biliary and pancreatic duct lesions between diffuse and focal AIP. The high expression of chronic pancreatitis characteristics is not observed in either group, which provides clues for the classification of AIP in clinical practice.

Evaluation of chemiluminescence immunoassay kit for detection of hepatitis D virus IgG antibody / 中华检验医学杂志

Objective:This study evaluates the performance of chemiluminescence assay, which is designed to detect Hepatitis D Virus (HDV) Immunoglobulin G (IgG) antibodies.Methods:A comparative analysis was conducted among chemiluminescence anti-HDV IgG reagent, the magnetic particle-based domestic reagent A and domestic reagent B, and the Robo Gene HDV RNA kit, using 1909 HBsAg-positive plasma samples. This comparison aimed to delineate clinical specificity and detection accuracy. The anti-HDV IgG reagent precision was assessed at three different concentration levels following the Clinical Laboratory Standards Institute EP5-A2 guidelines. The specificity of the assay was validated using 200 HAV IgM positive, 545 HBsAg-positive but anti-HDV IgG-negative, 350 anti HCV positive plasma samples and 200 healthy human blood samples. Additionally, a concordance study was conducted with 545 HBsAg-positive and 37 anti-HDV IgG-positive plasma samples, comparing the anti-HDV IgG reagent against reagent A.Results:1 909 HBsAg-positive plasma samples were tested using 3 anti HDV IgG reagent and 1 HDV RNA reagent, 19 samples were identified as anti-HDV IgG-positive. The anti-HDV IgG demonstrated superior accuracy and specificity. The assay exhibited excellent precision, with intra-assay coefficient of variation (CV) values ranging from 1.57% to 4.30%, and inter-assay CV values between 1.71% and 4.67% for detecting samples at high, medium, and low concentration levels. Concordance with Reagent A showed consistent results in both positive and negative detections.Conclusion:In this study, the anti-HDV IgG reagent (chemiluminescence method) displayed outstanding specificity in detecting clinical samples and exhibited a high conformity rate with commercialized reagents, making it potentially suitable for screening anti-HDV IgG in HBsAg-positive samples.

Antibody levels against varicella-zoster virus in healthy individuals aged 1 to 30 years in Songjiang District of Shanghai in 2022 / 上海预防医学

ObjectiveTo determine the antibody levels of varicella-zoster virus （VZV） in healthy individuals aged 1 to 30 years in Songjiang District of Shanghai， and to provide evidence for further development of varicella immunization strategies for healthy population. MethodsSix village committees were selected in Songjiang District through multi-stage stratified random sampling method from July to September 2022. Healthy individuals aged 1 to 30 years old in these villages were recruited and 3‒5 mL of venous blood was collected. Anti-VZV IgG antibody was tested by ELISA. Positive rate of anti-VZV IgG antibody and geometric mean concentration （GMC） were determined. ResultsA total of 315 healthy individuals aged （10.97±8.38） years were included in this study， of which 165 were males aged （11.00±8.52） years and 150 were females aged （10.96±8.25） years. Blood samples were collected and tested for anti-VZV IgG antibody， with the overall positive rate of 55.24% （174/315）. The GMC was calculated to be 99.73 mIU·mL-1， with the GMC in those tested positive for anti-VZV IgG antibody of 413.11 mIU·mL-1. The positive rate of anti-VZV IgG antibody and GMCs across age groups ranged between 13.33%‒86.67% and 16.36‒355.14 mIU·mL-1， respectively. The highest GMC was found in the group of 20‒30 years old （355.14 mIU·mL-1）， followed by 132.41 mIU·mL-1 in the group of 5‒ years old and 138.12 mIU·mL-1 in the group of 15‒ years old. Moreover， the positive rate was 38.47% in the group with 1-dose varicella vaccine and 63.49% in the group with 2 doses， while the GMC were 53.28 mIU·mL-1 and 130.79 mIU·mL-1 in these two groups， respectively. Multivariable analysis revealed that occupation was a risk factor associated with anti-VZV IgG antibody （OR=2.540）. Stratified analysis by varicella immunization history showed that among 2-dose vaccination group， time interval since the last dose was a protective factor （OR=0.315）. ConclusionsThe overall positive rate of anti-VZV IgG antibody in healthy population in Songjiang District of Shanghai remains low. The 2-dose varicella vaccination should be strengthened to improve the coverage in susceptible population.

CRX-527 as a candidate adjuvant in a recombinant BCG-based malaria vaccine

@#Abstract: Objective: To investigate the role of CRX-527, a Toll-like receptor 4 agonist, as the possible adjuvant for recombinant Mycobacterium bovis Bacillus Calmette-Guerin expressing merozoite surface protein 1C (BCG-MSP-1C). Methods: The mice were immunized with BCG and BCG-MSP- 1C in the presence and absence of CRX-527. The untreated mice (injected with PBS-T80 only) were the negative control. The ability of CRX-527 to enhance IgG and its subclasses, as well as IL-4 and IFN-γ production in the serum and spleen supernatant was evaluated using ELISA. Results: Mice immunized with BCG-MSP-1C exhibited the highest production of IgGs, IL-4 and IFN-γ after third immunization. In addition, CRX-527 further promoted the production of total IgG and IgG subclasses as well as IFN-γ and IL-4 in the serum and splenocytes of immunized mice. Conclusions: CRX-527 has the potential as an adjuvant candidate for the candidate vaccines. Further study is needed to verify appropriate dosage for immunization and its efficacy.

Immunogenicity evaluation of prototype and variant inactivated SARS-CoV-2 vaccines in rats / 中国生物制品学杂志

@#Objective To evaluate the immunogenicity of prototype strain,Beta strain,Gamma strain and Delta strain of SARS-CoV-2 inactivated vaccines in rats.Methods Five female Wistar rats were immunized with SARS-CoV-2 inactivated vaccines of prototype,Beta,Gamma and Delta strains through thigh muscle twice at an interval of 14 d,with an immunization dose of 3 μg virus protein/(0.5 mL per rat).Serum samples were collected and isolated by vein 14,28 and 42 d after the first immunization.The serum IgG antibody levels were detected by indirect ELISA,the titers of serum neutralizing antibody were measured by microneutralization,and the antigenic ratios of the serum neutralizing antibody titers were calculated to evaluate the antigenicity difference between different strains.Results at 14 d after the first immunization,IgG antibodies against four strains of virus were detected in all immunized serum samples.The levels of IgG antibodies increased by more than 10 times at 28 d compared with those at 14 d,and decreased slightly at 42 d.At 14 d after the first immunization,all the neutralizing antibodies against the four strains were positive in the serum of rats immunized with prototype strain or Delta strain vaccine;In the serum samples of rats immunized with Beta and Gamma strains,all the neutralizing antibodies against Beta and Gamma strains were positive,while some neutralizing antibodies against prototype or Delta strains were positive.At 28 d after the first immunization,the neutralizing antibodies in the immune serum of the four strains were positive,and the titers of neutralizing antibodies were significantly higher than those at 14 d;The neutralizing antibody titers were slightly lower at 42 d after the first immunization than 28 d.There was small difference in the antigenicity between Beta and Gamma,prototype and Gamma,but significant difference in the antigenicity between prototype and Beta strains.Conclusion The prototype strain,Beta strain,Gamma strain and Delta strain of SARS-CoV-2 inactivated vaccines can stimulate rats to produce neutralizing antibodies with high titer,while the immunogenicity has difference.

Expression of Gn protein of severe fever with thrombocytopenia syndrome virus and its immunogenicity / 中国生物制品学杂志

@#Objective To express the Gn protein of severe fever with thrombocytopenia syndrome virus(SFTSV) through adeno-associated virus 9(AAV9) expression system and evaluate its immunogenicity.Methods SFTSV Gn gene was inserted into viral vector pAAV-CMV-FH and the recombinant plasmid was transfected into HEK293T cells to obtain recombinant virus AAV9-Gn.The expression of Gn protein was determined by immunofluorescence and Western blot.Eighteen fernale BALB/c mice were randomly divided into three groups:Mock group(serum-free DMEM),AAV9-GFP group(1 × 10~(11) vg) and AAV9-Gn group(1 × 10~(11) vg),all of which were injected intramuscularly into the right hind limb at a dose of 100 μL per mouse.The body mass,diet,behavior and mental state of mice in each group were monitored continuously for 21 d,and the change rate of body mass was calculated;At 2,4,8 and 16 weeks after immunization,the levels of SFTSV neutralizing antibody in serum of mice in each group were detected by fluorescent reduction neutralization test(FRNT),and the levels of specific IgGl and IgG2a in serum of mice in AAV9-Gn group were detected by ELISA.Results After incubation with specific antibody,Vero cells transfected with AAV9-Gn showed specific green fluorescence under fluorescence microscope,and had specific binding to mouse anti-SFTSV Gn monoclonal antibody,and the specific binding bands were found at a relative molecular mass of about 61 000.The body mass of the three groups showed an increasing trend,there was no significant difference between the three groups(F=0.158—2.621,P> 0.05),and the diet,behavior and mental state were normal.At 2,4,8 and 16 weeks after immunization,the titer of SFTSV neutralizing antibody in serum of mice in AAV9-Gn group was significantly higher than that of Mock group and AAV9-GFP group(H=13.332—14.538,each P <0.001),and the titer peak appeared at 8 weeks;The level of specific IgG1 in serum of mice was significantly higher than that of IgG2a(F=4.373—12.975,each P <0.05) at different time points.Conclusion SFTSV Gn protein can be expressed correctly through AAV9 expression system,and has low toxicity to mice with good immunogenicity,which is expected to be a candidate component of SFTSV vaccine.

Serological analysis of severe acute respiratory syndrome coronavirus 2-specific immunoglobulin G antibodies level in Henan Province / 中华传染病杂志

Objective:To analyze the specific immunoglobulin G (IgG) antibodies level in the population after the coronavirus disease 2019 (COVID-19) pandemic in Henan Province.Methods:A total of 5 178 peripheral venous blood samples were collected from 10 districts (counties) in Henan Province according to the national seroepidemiological survey program for COVID-19, and the method of cluster random sampling was adopted from March 6 to 15, 2023. Descriptive analysis was used for the basic data, history of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) vaccination, SARS-CoV-2 infection of the respondents. The specific IgG antibody of SARS-CoV-2 was detected using chemiluminescence method. Statistical analysis was performed by using rank sum test, Kruskal Wallis test, and Dunn′s test.Results:The overall positive rate of SARS-CoV-2-specific IgG antibody was 83.35%(4 316/5 178). There were statistically significant differences in the specific IgG antibodies against SARS-CoV-2 produced by people of different sexes, different ages, infected or not, vaccinated or not, and vaccinated with different doses of SARS-CoV-2 vaccine ( Z=3.60, H=195.32, Z=6.10, 18.08, H=382.70, respectively, all P<0.001). The specific IgG antibodies produced by unvaccinated+ uninfected group, unvaccinated+ infected group, vaccinated+ uninfected group, and vaccinated+ infected group were 3.54(0.98, 11.00), 60.65(2.33, 84.80), 133.00(59.80, 173.00), and 142.00(98.30, 176.00), respectively. And the difference was statistically significant( H=354.62, P<0.001). The specific IgG antibodies of uninfected people increased with the increase of inoculum times( H=287.00 and 98.48, both P<0.001). The specific IgG antibodies of people who were not infected with SARS-CoV-2 in the groups of whose interval from the last inoculation of SARS-CoV-2 vaccine to blood collection was less than three months, three to six months and more than six months were 171.86(156.04, 196.57), 71.71(17.08, 110.38) and 132.14(57.59, 172.25), respectively, and the difference was statistically significant ( H=19.93, P<0.001). Among them, the absolute difference between the less than three months group and the three to six months group was statistically significant ( Z=3.67, P<0.001), and the absolute difference between the less than three months group and the more than six months group was statistically significant ( Z=3.47, P<0.001). The specific IgG antibodies level in the less than three months group was the highest. Conclusions:There is a certain correlation between the number of SARS-CoV-2 vaccine doses and the specific IgG antibodies level in uninfected people. The specific IgG antibodies could maintain a high level for three months after immunization.

Chinese guideline for diagnosis and treatment of autoimmune pancreatitis （Shanghai， 2023） / 临床肝胆病杂志

Autoimmune pancreatitis （AIP） is an immune⁃mediated， special type of chronic pancreatitis， which can involve multiple organs. The clinical manifestation of AIP is complex and varied， making the diagnosis and treatment challenging. With reference to the latest guidelines and studies from both domestic and international sources， this guideline comprises 20 recommendations regarding the diagnosis， treatment， follow⁃up， and prognosis of AIP. The aim of this guideline is to promote the care capability and improve the outcome of patients with AIP in China.

Development and verification of a reporter gene determination assay for ADCP potency of IgG2 anti-EGFR monoclonal antibody based on DOE / 中国生物制品学杂志

@#Objective To develop and verify a reporter gene assay for the determination of antibody dependent cellular phagocytosis(ADCP)potency of Ig G2 monoclonal antibody(m Ab)against epidermal growth factor receptor(EGFR)by combining Design of Experiment(DOE)and one factor at a time(OFAT).Methods The Jurkat/NFAT-Re/FcγRⅡa stably transformed cell line was used as effector cells,while the A431 cell line as the target cells.The JMP software was used to optimize the seven key factors in the experiment by combining DOE and OFAT analysis,while the ratio of upper and lower asymptotes(D/A)was used as the statistic,and the reporter gene method was developed to evaluate the ADCP potency of Ig G2 anti-EGFR m Ab.The method was verified according to the general chapter<9401>of Chinese Pharmaco-poeia（Ⅲ/Ⅳvolume,2020 edition)and used to determine the biological potency of Ig G2 anti-EGFR m Ab injection.Results After three rounds of experiments,the reporter gene method to evaluate the ADCP potency of Ig G2 anti-EGFR m Ab was developed.The method showed a dose-response relationship and was consistent with the four-parameter regression equa-tion y=(A-D）/[1+(x/C)~B]+D.The range of seven key conditions was determined:the density of effector cells was(1.25-3.75）×10~4 cells/well,the density ratio of effector cells to target cells was 1.0-2.0,the incubation time of target cells was 20-40 min,the incubation time of administration was 15-30 min,the total time was 5.5-6.5 h,and the color time was 5-30 min with luciferase detection system(Bright-Glo)as the color agent.The method had good specificity.Six independent tests were run for the five potency levels,with the correlation coefficient r of 0.994 5 and the linear regression equation slope of 1.02.The relative potency of five potency levels respectively was(62.15±1.38)%,(78.53±2.82)%,(99.12±3.95)%,(123.27±4.59)%and(155.22±7.04)%,the range of relative biases was-2.9%-0.2%,and the range of generalized cross-validation(GCV)was 2.2%-4.6%.The method had good linearity,relative accuracy and precision in the range of 64%-156%.The mean value of the potency of IgG2 anti-EGFR m Ab in three tests was(101.5±2.8)%.Conclusion The reporter gene assay developed in this study can be used to evaluate the ADCP potency of IgG2 anti-EGFR mAb

Application of monocyte monolayer assay on hemolytic disease of fetus and newborn caused by IgG anti-M / 中国输血杂志

【Objective】 To elucidate the prediction ability of monocyte monolayer assay(MMA) used in hemolytic disease of fetus and newborn(HDFN) caused by IgG anti-M. 【Methods】 Plasma from eight pregnant women containing IgG anti-M were collected, and were divided into two groups(4 cases with HDFN, with severe clinical symptoms such as fetal hydrops, and 4 cases without HDFN) according to the clinical outcomes. M antigen positive cells were sensitized with dithiothreitol(DTT) treated plasma from eight pregnant women respectively. MMA was performed by coincubation with monocytes and sensitized M cells, along with negative and positive control set up. T-test was conducted to compare the difference in phagocytic efficiency between two groups. 【Results】 The phagocytic efficiency in group with HDFN were 15.37%, 13.05%, 9.17% and 24.50% respectively, with the mean value of 15.52%, while the group without HDFN were 8.74%, 11.07%, 5.12% and 6.23% respectively, with the mean value of 7.79%.There was no significant difference in phagocytic efficiency between two groups(P>0.05). The mean values of both groups were not significantly different from the negative control(P>0.05), but both were significantly lower than positive control(P<0.05). 【Conclusion】 The low phagocytic efficiency couldn’t convince that the MMA is an effective predictor for the HDFN caused by IgG anti-M, indicating that another mechanism might be responsible for it rather than monocyte phagocytosis. The assessment of the peak systolic velocity in middle cerebral artery of the fetal should be considered in the management for pregnant women who produce IgG anti-M to estimate the situation of fetal anemia.

Changes in serum immunoglobulin levels in children with thalassemia who undergo repeated blood transfusions and their correlation with delayed hemolytic transfusion reactions / 中国输血杂志

【Objective】 To study the changes in serum immunoglobulin levels in children with thalassemia who undergo repeated blood transfusions and explore their correlation with delayed hemolytic transfusion reactions(DHTR). 【Methods】 Serum samples from children with thalassemia who received blood transfusion treatment from June 2022 to April 2023 (observation group) and healthy children who underwent physical examination (control group) in our hospital were collected. The levels of serum immunoglobulins (IgG subtype, IgM, IgA, IgE and IgD) were detected using flow cytometry CBA multi-factor quantitative detection technology, and the differences between the two groups were compared. The children were divided into 4 groups according to different transfusion numbers: ≤10 numbers, 11-30 numbers, 31-50 numbers and >50 numbers, and the differences between different blood transfusion numbers and serum immunoglobulin levels in each group were compared using one-way analysis of variance (ANOVA). Children with thalassemia with DHTR were in the hemolysis group, and children with thalassemia who did not experience DHTR were in the non-hemolysis group. The changes in serum immunoglobulins (IgG subtypes, IgM, IgA, IgE and IgD) between the two groups were compared to explore the correlation between serum immunoglobulins in thalassemia children with repeated transfusion and DHTR. 【Results】 The levels of IgG1, IgG3, IgG4 and IgA in the observation group were significantly higher than those in the control group, with the increase of(2.07±2.12), (0.67±2.03), (0.30±0.37)and(6.04±11.40)mg/mL, respectively, while the level of IgD in observation group was significantly lower than that in the control group, with a decrease of(0.03±0.01)mg/mL, P0.05). IgG1 and IgG4 both significantly increased with the number of blood transfusions.The IgG1 in the 4 groups increased sequentially as(0.30±0.62), (0.41±0.51)and(3.60±3.48)mg/mL, and IgG4 increased sequentially as (0.12±0.13), (0.22±0.07) and (0.21±0.38)mg/mL. IgG2, IgM and IgD showed a significant decrease, with IgG 2, IgM, and IgD in four groups decreased as(0.91±1.50), (0.14±0.10)and(0.05±0.05)mg/mL, respectively, showing significant differences with the number of blood transfusions(P0.05). IgG1, IgG3 and IgG4 in the hemolysis group were significantly higher than those in the non-hemolysis group, with an increase of (4.44±3.41), (0.73±1.26)and(0.52±0.40), respectively(P0.05). 【Conclusion】 The serum immunoglobulin levels of children with thalassemia who undergo repeated blood transfusions are abnormal. There are differences in correlation between the number of blood transfusions and serum immunoglobulin levels among children with thalassemia who undergo repeated blood transfusions. The relevant serum immunoglobulins for DHTR in children with thalassemia who undergo repeated blood transfusions are IgG1, IgG3 and IgG4.

Application of anti-idiotypic antibodies in antibody screening and crossmatch tests of patients treated with CD47 monoclonal antibody / 中国输血杂志

【Objective】 To perform pre-transfusion examination and major crossmatch test using CD47 anti-idiotypic antibody (CD47 AID) (method 1) and reagent lack of anti-IgG4 anti-human globulin(method 2) in patients treated with CD47 monoclonal antibodies, and evaluate the feasibility of method 1 by comparing the transfusion efficacy of patients after cross matching with two methods. 【Methods】 Post-drug samples were collected from 18 clinical subjects treated with CD47 monoclonal antibody in our hospital. Antibody screening and major crossmatch test were performed using method 1 and method 2, and the difference of ΔHb (post-transfusion Hb minus pre-transfusion Hb) was compared after transfusion. The differences in ΔHb after transfusion were analyzed between the test group using method 1 and the control group without CD47 monoclonal antibody using ordinary microcolumn gel method. 【Results】 There was no significant difference in ΔHb between the test group using method 1 and test group using method 2 (8.40±0.71 vs 7.36±0.94, P>0.05). No significant difference was noticed in ΔHb between the test group using method 1 and the control group without CD47 monoclonal antibody (8.40±0.71 vs 6.59±0.77, P>0.05). 【Conclusion】 In the test group, major crossmatch test with method 1 has the same transfusion efficacy as the test with method 2. Method 1 is simple and easy to operate, and the results are objective and accurate. It is recommended to use method 1 for pre-transfusion antibody screening and major crossmatch tests for patients using CD47 monoclonal antibody.

Evaluation of different cut-off points for IgG avidity and IgM in the diagnosis of acute toxoplasmosis in pregnant women participating in a congenital toxoplasmosis screening program

ABSTRACT The main social impact of toxoplasmosis stems from its ability to be vertically transmitted. Postnatally acquired infection is generally asymptomatic in approximately 70-90% of cases, making diagnosis often dependent on laboratory tests using serological methods to search for anti-T. gondii antibodies. This study aimed to investigate the ability of the VIDAS TOXO IgG avidity and VIDAS TOXO IgM assays to confirm recent toxoplasmosis. In total, 341 pregnant women with suspected acute toxoplasmosis were systematically monitored in the Program for Control of Congenital Toxoplasmosis in Minas Gerais State, Brazil. We conducted an observational analytical-descriptive cross-sectional study and grouped according to clinical and laboratory criteria as having acute or chronic toxoplasmosis. The VIDAS TOXO IgG avidity and VIDAS TOXO IgM assays were evaluated to investigate the capacity to identify acute infection. IgG avidity showed good performance in identifying acute toxoplasmosis when the IgG avidity index was lower than or equal to 0.1. Values greater than or equal to 3.16 according to the TOXO IgM kit were associated with a greater chance of acute infection. These results may contribute to a more adequate diagnosis of acute gestational toxoplasmosis and, consequently, the avoidance of inadequate or unnecessary treatments.

Allergenic response induced by Pterobothrium crassicolle (Cestoda: Trypanorhyncha) extracts in murine model / Resposta alergênica induzida por extratos de Pterobothrium crassicolle (Cestoda: Trypanorhyncha) em modelo murino

Abstract The aim of this study was to determine the allergenic activity of components present in crude extracts of Pterobothrium crassicolle plerocerci (CPE) and blastocysts (CBE) obtained from Micropogonias furnieri in a murine model. Two groups of seven animals each received 50 µg of CPE or CBE on days 1, 35 and 120. Serum samples were tested by ELISA and Immunoblotting. Specific IgG and IgE levels were detected by ELISA, showing specific humoral responses for the primary immunization for both immunoglobulins and continuously growing titers for IgE. Positive Passive Cutaneous Anaphylaxis tests in rats sensitized with anti-CBE sera and tested by CBE, showed biologically, the allergenic activity of the extracts. The CPE and CBE showed some different recognition regions but both experimental groups recognized all regions of the extracts when tested for cross reactions, showing that CPE and CBE could share antigenic recognition sites.

Resumo O objetivo deste estudo foi determinar a atividade alergênica de componentes presentes em extratos crus de plerocercos (CPE) e de blastocistos de Pterobothrium crassicolle (CBE), obtidos de Micropogonias furnieri, em modelo murino. Dois grupos de sete animais receberam cada um 50 µg de CPE ou CBE nos dias 1, 35 e 120. As amostras de soro foram testadas por ELISA e Imunoblot. Os níveis específicos de IgG e IgE foram detectados por ELISA, mostrando respostas humorais específicas para a imunização primária para ambas as imunoglobulinas e títulos crescentes de IgE. Testes positivos de Anafilaxia Cutânea Passiva em ratos sensibilizados com soros anti-CBE e testados por CBE, demonstraram biologicamente, a atividade alergênica dos extratos. O CPE e o CBE evidenciaram algumas regiões de reconhecimento diferentes, mas ambos os grupos experimentais reconheceram todas as regiões dos extratos, quando testados para reações cruzadas, mostrando que o CPE e o CBE poderiam compartilhar locais de reconhecimento antigênico.