The role of pathogen infections in the development and progression of autoimmune liver diseases / 临床肝胆病杂志

The etiology and pathogenesis of autoimmune liver diseases has always been a hot area of research. Pathogen infections can elicit an autoimmune response and often become the key pathogenic factor of immune diseases. Based on the literature data and the author's clinical experience, this review will briefly introduce the role and influence of pathogen infections in the development and progression of autoimmune liver diseases from the aspects such as molecular mimicry mechanism, in order to further understand the pathogenesis of autoimmune liver diseases.

Research advances in autoantibodies in autoimmune hepatitis / 临床肝胆病杂志

Autoantibodies have an important value in the diagnosis, typing, and differential diagnosis of autoimmune hepatitis (AIH); however, the autoantibodies of AIH lack specificity and have various types and inconsistent results, which brings confusions to clinical diagnosis. With reference to the literature data and the author’s clinical experience, this article reviews the application value and research advances in the main autoantibodies in AIH and introduces the recommendations for autoantibody detection methods in related guidelines.

Analysis of the autoantibodies characteristics of 77 anti-soluble liver antigen positive patients with liver diseases / 中华检验医学杂志

Objective@#To understand the characteristics and clinical significance of anti-soluble liver antigen antibody (anti-SLA) in patients with liver diseases.@*Methods@#Serum samples from seventy-seven patients with anti-SLA were collected from Beijing You'An Hospital during the period between January 2010 and December 2018. Anti-SLA, anti-liver cytosol type 1 antibody (anti-LC1), anti-glycoprotein 210 antibody(anti-gp210) and anti-nuclear body protein sp100 antibody(anti-sp100) were detected by immunoblotting; indirect immunofluorescence assay used for detecting anti-nuclear antibody (ANA), anti-mitochondrial antibody (AMA), anti-smooth muscle antibody (SMA), and anti-liver kidney microsome antibody (anti-LKM). One-way analysis of variance was used to compare the ages of different anti-SLA groups. The non-parametric rank sum test was used to compare the liver function indexes and immunoglobulins in different intensity groups of anti-SLA. P<0.05 was considered statistically significant. Further comparisons were made between the two groups, the correction level α′=0.008 3, P<0.008 3 was considered statistically significant.@*Results@#The average age of 77 anti-SLA positive patients was (52.50±1.25) years old, 70 females (90.9%) and 7 males (9.1%). 80.5% of anti-SLA-positive patients (62/77 cases) were strongly positive at the time of initial diagnosis (+++ to ++++).The Alanine aminotransferase (ALT) level in the SLA++ group was higher than that in the SLA++++ group (232.7 U/L vs 65.6 U/L,χ2=7.751,P=0.005) and the immunoglobulin M(IgM) level in the SLA+++ group was lower than that in the SLA++++ group (1 270 mg/L vs 2 270 mg/L,χ2=8.337,P=0.004).There was no significant difference in age, other liver function and immunological indicators among the different groups.Seventy cases (90.9%) were both anti-SLA and ANA positive, 13 cases (16.9%) were positive with SMA, and none positive with anti-LKM and anti-LC1. Among anti-SLA positive patients, 58 cases were diagnosed with autoimmune hepatitis (AIH), 12 were AIH/primary biliary cholangitis (PBC) overlap syndrome (OS), 2 were drug-induced liver injury, 2 were chronic hepatitis B, and 3 were hepatitis A, hepatitis E and acquired immune deficiency syndrome (AIDS) with liver injury, respectively.Cases of AIH and AIH/PBC OS accounted for 90.9% (70/77 cases) of anti-SLA-positive patients, and 5 of 7 patients diagnosed with non-AIH (and OS) had elevated IgG, showing AIH feature.92.3% (12/13 cases) of anti-SLA with high titers of AMA were diagnosed as AIH/PBC overlap syndrome. Of the 77 anti-SLA-positive patients, 28 (36.4%) had advanced or end-stage liver disease, including decompensated cirrhosis (22 cases), chronic acute liver failure (4 cases), and liver transplantation (1 case) and death from liver failure (1 case).@*Conclusions@#Anti-SLA has high diagnostic specificity for AIH;anti-SLA positive in patients with PBC should be an important biomarker for the diagnosis of AIH/PBC overlap syndrome.

An analysis of the serological characteristics of anti-mitochondrial M2 subtype in patients with drug-induced liver injury and primary biliary cholangitis / 中华肝脏病杂志

Objective@#To analyze the serological characteristics of anti-mitochondrial antibody M2 subtype (AMA-M2) in patients with drug-induced liver injury (DILI) and primary biliary cholangitis (PBC), in order to provide reference for clinical differential diagnosis.@*Methods@#Laboratory data of 2802 DILI cases who visited the hospital between January 2011 and December 2017 were retrospectively collected. AMA-M2 positive patients were analyzed with respect to laboratorical findings, and serum data of 120 patients with primary biliary cholangitis (PBC) at the same period was taken as a control. A chi-square test was used for group comparisons. One-way ANOVA and rank sum tests was used for ALT, AST, ALP, GGT and three groups of immunoglobulin M.@*Results@#Among 2802 DILI patients, AMA-M2 positive rate was 5.1% (144/2 802), 77.1% (111/144) was DILI alone, 22.2% (32/144) was DILI with PBC, and 0.7% (1/144) was DILI with Sjogren's syndrome. An AMA-M2 level in DILI alone group was mostly mild and moderate than the PBC group and the DILI combined with the PBC group. There was significant difference between the two groups (P < 0.05).There was no significant difference in AMA-M2 levels between DILI group combined with PBC group and PBC group (P > 0.05). ALT and AST levels of DILI alone group and DILI combined with PBC were (585.92 ± 653.04) U/L, (501.45 ± 512.67) U/L and (373.47 ± 502.60) U/L, (335.97 ± 513.96) U/L, respectively, which were significantly higher than PBC group [(106.33 + 134.08) U/L, (112.59 + 152.20) U/L]. There were statistically significant differences between the two groups (P < 0.05).The ALP level of DILI alone group was (152.58 + 81.46) U/L, which was lower than PBC group (237.86 + 215.09). The difference was statistically significant (P < 0.05). The level of immunoglobulin M in the DILI alone group was (1.76 ± 1.16) g/L, which was lower than PBC group (4.74 ± 5.74) g/L and the DILI combined with the PBC group (3.31 ± 1.68) g/L. There was significant difference between the two groups. During follow-up, 2.7% of patients with DILI had cirrhosis, 42.3% had lower AMA-M2 titer, 14.4% had lower AMA-M2 titer, 13.5% had higher AMA-M2 titer and five cases developed PBC.@*Conclusion@#AMA-M2 is not only positive in patients with PBC, but also low-to-medium or even high-level AMA-M2 may be detected in DILI patients. For AMA-M2-positive DILI patients, it is necessary to identify whether they are associated with PBC. Secondly, the levels of ALT, AST and ALP should be analyzed, and the patients should be on regular follow up for early and timely detection of drug-induced PBC.

Detection and Clinical Significance of Autoantibodies in Autoimmune Liver Diseases / 胃肠病学

Autoimmune liver diseases (AILD)are a group of chronic liver disease induced by autoimmune disorders, one or multiple autoantibodies usually present in AILD patients,and detection of autoantibodies has become an important tool for the diagnosis and differential diagnosis of AILD. For some reasons,such as the differences of physicians' understanding for autoantibodies,the diversity of methodology and combination of items detected in different hospitals, there are still some puzzles in interpretation of results of autoantibodies. In this article,the detection of autoantibodies in AILD is discussed along with their clinical significance to help the clinical interpretation of the significance of autoantibodies.

The comparison of different immunoassays for the detection on autoantibodies specific to primary biliary cholangitis / 中华检验医学杂志

Objective To compare the test performance of different immunoassays for the detection on autoantibodies specific to primary biliary cholangitis,including anti-mitochondrial type 2 antibody(AMA-M2),anti-glycoprotein 210(anti-gp210)and anti-nuclear body protein sp100(anti-sp100).Methods Serum samples from Primary Biliary Cholangitis(PBC, n=91), liver disease control(including viral hepatitis,autoimmune hepatitis and liver cirrhosis,n=67)and healthy individual(n=40)were collected from Beijing Youan Hospital during the period between April 2014 and April 2017.All samples were tested with chemiluminescent immunoassay(CLIA)and enzyme linked immunosorbent assay(ELISA)for AMA-M2, meanwhile the detection on anti-gp210 and anti-sp100 were compared between CLIA and Line Immunoassay(LIA).The Kappa coefficient were used to measure the level of qualitative agreement between different assays.The diagnostic accuracy of AMA-M2 detected with CLIA and ELISA were compared by receiver operating characteristic curve(ROC).Results The overall qualitative agreement between CLIA and ELISA for the detection to AMA-M2 is 88.4%(Kappa =0.765, P<0.01).Excellent qualitative agreement between CLIA and LIA for the detection to anti-gp210 and anti-sp100 was also found with overall agreement as 96.5%(Kappa=0.852,P<0.01)and 98%(Kappa=0.884,P<0.01), respectively.The ROC analysis also showed similar area under the curve(AUC)for CLIA(0.965, P<0.01)and ELISA (0.928,P<0.01)on detection to AMA-M2.Conclusions CLIA and ELISA showed excellent agreement for the detection to AMA-M2.High qualitative agreement between CLIA and LIA was also found when testing anti-gp210 and anti-sp100.

Research advances in immune repertoire and high-throughput sequencing in primary biliary cholangitis / 临床肝胆病杂志

Primary biliary cholangitis (PBC) is an autoimmune liver disease with unclear pathogenesis.The amino acid composition and sequence in the complementarity-determining region 3 of T cell receptor (TCR) and B cell receptor (BCR) are highly diverse, which forms a large antigen recognition receptor repertoire, i.e., immune repertoire.In recent years, second-generation sequencing techniques combined with multiplex PCR or amplicon rescue multiplex PCR have been used to study the features of immune repertoire in PBC patients, and it has been found that PBC patients have clonal expansion of specific CD4+ T lymphocytes, clonal diversity of B lymphocytes, somatic hypermutation, and reduction in class switch, as well as increase in clonal diversity after treatment with ursodeoxycholic acid.These findings need to be confirmed by large-scale in vivo and in vitro studies and different immune repertoire research strategies.

Clinical value of anti-liver/kidney microsomal-1 antibody in patients with liver disease / 中华肝脏病杂志

Objective@#To investigate the clinical and laboratory features of patients with liver disease and positive anti-liver/kidney microsomal-1 (anti-LKM-1) antibody, and to provide a reference for clinical diagnosis and differential diagnosis.@*Methods@#The clinical data of patients with positive anti-LKM-1 antibody who were treated in our hospital from 2006 to 2016 were collected, and clinical and laboratory features were analyzed and compared. An analysis was also performed for special cases.@*Results@#The measurement of related autoantibodies was performed for about 100 thousand case-times, and 15 patients were found to have positive anti-LKM-1 antibody. Among the 15 patients, 7 were diagnosed with type 2 autoimmune hepatitis (AIH) with an age of 11.0 ± 9.0 years and were all adolescents with acute onset; 8 were diagnosed with hepatitis C with an age of 51.5 ± 9.0 years, among whom 7 were middle-aged patients and 1 was a child aged 12 years, and all of them had an insidious onset. Compared with the patients with hepatitis C, the AIH patients had significantly higher levels of alanine aminotransferase (1 003.9 ± 904.3 U/L vs 57.0 ± 84.1 U/L, P < 0.05), aspartate aminotransferase (410.7 ± 660.3 U/L vs 34.9 ± 42.9 U/L, P < 0.05), and total bilirubin (98.0 ± 191.0 μmol/L vs 15.4 ± 6.0 μmol/L, P < 0.05). There was a reduction in immunoglobulin G after the treatment with immunosuppressant, compared with the baseline. Of all 8 patients with hepatitis C, 6 received antiviral therapy with interferon and ribavirin, and 5 out of them achieved complete response, among whom 4 had a reduction in the level of anti-LKM-1 antibody after treatment; however, a 12-year-old child developed liver failure after interferon treatment and died eventually.@*Conclusion@#Positive anti-LKM-1 antibody is commonly seen in patients with type 2 AIH or hepatitis C, but there are differences between these two groups of patients in terms of age, disease onset, liver function, and the level of anti-LKM-1 antibody. The hepatitis C patients with a confirmed diagnosis and exclusion of autoimmune hepatitis can achieve good response to interferon under close monitoring, even if anti-LKM-1 antibody is positive. As for adolescent patients with hepatitis C and positive anti-LKM-1 antibody, the possibility of AIH should be excluded.

Characteristics of IgH-CDR3 repertoire of peripheral B cells in a patient with primary biliary cholangitis: a preliminary study using high-throughput sequencing / 中华肝脏病杂志

Objective@#To analyze the characteristics of immunoglobulin heavy chain complementarity-determining region (IgH-CDR3) repertoire of peripheral B cells in a patient with primary biliary cholangitis (PBC) and to investigate the diversity of the immune system.@*Methods@#Arm-PCR was used to amplify the IgH-CDR3 region of circulating B cells isolated from a PBC patient, and high-throughput sequencing was used to analyze the amplified product. The characteristics of immune repertoire were analyzed by bioinformatics.@*Results@#In total, 329219 sequence reads were generated from the sample, with 325540 total CDR3 sequences and 72774 distinct CDR3 sequences, and the D50 of IGH-CDR3 was 7.7. The dominant CDR3 length of the sample was 45 nt (9.6%); the N addition with the highest frequency ranged from 13 to 14 nt (5.25%); the J trimming with the highest frequency was 0 nt (12.7%); the three most frequent V alleles were V4-59 (9.5%), V3-23 (8.1%), and V1-69 (6.4%).@*Conclusion@#The diversity of IgH-CDR3 repertoire is relatively low in this patient with PBC, with several B-cell clonal expansions. The specificity needs to be further verified after increasing the sample size.

How to understand the clinical significance of autoantibodies in primary biliary cholangitis / 中华肝脏病杂志

Autoantibodies are important indicators for the diagnosis of primary biliary cholangitis (PBC). The autoantibodies in PBC patients are mainly antimitochondrial antibodies (AMAs) and antinuclear antibodies (ANAs). AMAs are one of the diagnostic indices of PBC. PBC-specific ANAs (nuclear dots or nuclear envelope, anti-sp100, and anti-gp210) have a high specificity in the diagnosis of AMA-negative PBC. This article reviews the clinical significance of these autoantibodies and analyzes some misconceptions about the clinical diagnosis of AMA-negative PBC and PBC-AIH overlap syndrome.

The relationship between plasma soluble CD14 level and disease progression in patients with acute phase of acquired immunodeficiency syndrome / 中华传染病杂志

Objective To investigate the correlation between plasma soluble CD14 (sCD14)level and disease progression in patients with acute phase of acquired immunodeficiency syndrome (AIDS). Methods Forty-one human immunodeficiency virus (HIV)-infected patients were followed up from June 2007 to June 2010 in Beijing You′an Hospital,including 20 patients with CD4 + T lymphocyte counts more than 350/μL,and 21 less than 350/μL after 3 years of HIV infection.Twenty healthy blood donors were recruited as controls.Enzyme-linked immunosorbent assay (ELISA)was employed to test the plasma sCD14 level of healthy controls and patients infected with HIV at 1 -30 d,31 -90 d,91 - 180 d and 181 -360 d.Student t test was used to compare the means between two groups.ANOVA analysis was used to compare the means among more than two groups.Results The mean plasma sCD14 level in control group was (1 654±904)μg/L.Three years after HIV infection,the sCD14 level of patients with CD4 + T lymphocyte counts less than 350/μL group was (4 214±2 635)μg/L,which was higher than that of patients with CD4 + T lymphocyte counts more than 350/μL ([2 275 ±1 457 ]μg/L).The difference was statistically significant(t=-5 .41 ,P <0.01).The plasma sCD14 level in patients infected with HIV 181 -360 d was significantly higher than that in patients infected with HIV 1 - 30 days ([4 485 ± 2 779]μg/L vs [2 577 ±1 635 ]μg/L;t = -3.39,P <0.05 ).The plasma sCD14 level was positively correlated with HIV viral load (r =0.35,P =0.000 1 ),and negatively correlated with CD4 + T lymphocyte counts (r=-0.28,P =0.001 ).Conclusions The plasma sCD14 level in patients with acute phase of HIV infection is higher than that of healthy people,which increases with prolonged HIV infection.Plasma sCD14 level in of HIV infection acute phase may be closely related to HIV/AIDS progression.

Clinical symptoms and courses among men who have sex with men with acute human immunodeficiency virus infection / 中华传染病杂志

Objective To describe the clinical characteristics,CD4+ and CD8+ T cell counts as well as human immunodeficiency virus (HIV) RNA of acute HIV infection in men who have sex with men,and their correlations with the disease progression.Methods One hundred cases of acute HIV infection were followed up.Nuclear acid sequence-based amplification (NASBA) was used for plasma HIV RNA screening.Flow cytometry was served to test the CD4+ and CD8+ T cell counts.Hepatitis B surface antigen (HBsAg) and anti-hepatits C virus (HCV) antibody were detected using enzyme-linked immunosorbent assay.Rapid plasma reagin was applied to screen for Treponema pallidum antibody.Antibody positive specimens were tested with Treponema pallidum particle assay for validation.The positive results were identified as infection.Results Ninety-six cases were aged between 20 and 50 years old.Among 100 cases,9 were HBsAg-positive; 4 were anti-HCV positive; 40 were co-infected with syphilis.During the follow-up period,the median CD4+ T cell counts in the 1st and 3rd month were 510/μL and 499/μL,respectively.The median HIV RNA in 1st,3rd,6th and 24th month were 4.37,4.00,4.31 and 4.43 lg copy/mL,respectively.CD8+ T cell counts did not show significant change during the study.Among 38 rapid progressors,the initial mean CD4+ T cell counts was (358.0± 134.6)/μL,which was significantly lower than that of the non-rapid progressors with a mean CD4+ T cell counts of (559.2±203.4)/μL.Meanwhile,the initial mean HIV RNA of the rapid progressors was 4.71 lg copy/ mL,while that of the non rapid progressor was 4.18 lg copy/mL.The initial CD8+ T cell counts of the rapid progressors and non rapid progressors were 1 250.1/μL and 1 247.2/μL,respectively.Conclusions Acute HIV-1 infected men who have sex with men tend to be young.The initial CD4+ T cell counts and HIV RNA during acute infection could be used to predict the disease progress.

Research progress in role of B cell immunity in primary biliary cirrhosis / 临床肝胆病杂志

Primary biliary cirrhosis (PBC)is characterized by injury of small intrahepatic bile ducts,but its mechanism,especially the role of B cell immunity in this disease,remains unclear.The possible role of B cell immunity in the pathogenesis of PBC is reviewed from the fol-lowing aspects:distribution of B cells and plasma cells in the liver tissues of PBC patients,clearance of B cells,mechanism of abnormal in-crease in serum IgM and its sources among PBC patients,mechanism of production of antimitochondrial antibodies and their possible func-tions in bile duct injury,and typical autoantibodies in PBC.B cell immunity is considered to play a significant role in the pathogenesis of PBC,but there are still many key problems to be clarified.

Bw4 motif expressed on HLA-B antigen affects HIV-1 specific T cell responses induced in patients with acute HIV-1 infection / 中华微生物学和免疫学杂志

Objective To investigate whether Bw4 motif expressed on HLA-B affects Gag-specific T cell responses in patients with acute HIV-1 infection.Methods Sequence specific primer polymerase chain reaction ( SSP-PCR) was performed for human leukocyte antigen ( HLA) typing.Peripheral blood mononuclear cells ( PBMCs) from 36 patients with six months of acute HIV-1 infection were stimulated with HIV-1 CRF01_A/E Gag peptides to detect the HIV-1 specific T cell responses by using ELISPOT assay. Results (1) The set point viral load of 18 patients carrying no Bw4 motif on HLA-B was 4.49±0.56 which was higher than that in other 18 patients carrying 1-2 Bw4 motif(s) on HLA-B (3.78±0.75) (P=0.005). (2) T cells from 26 out of 36 patients with acute HIV-1 infection responded to P24 peptides pool including 15 patients carrying no Bw4 motif on HLA-B and 11 patients carrying 1-2 Bw4 motif( s) on HLA-B, but no significant difference was observed between them (P>0.05).The magnitude of P24-specific T cell responses induced in patients carrying no Bw4 motif on HLA-B was (1317.8 ±1238.0) SFC/106 PBMCs which was greater than that induced in patients carrying 1-2 Bw4 motif(s) on HLA-B [(549.9±778.5) SFC/106 PBMCs] ( P=0.032) .The breadth of T cell responses to P24 peptides was 2(0-5) in patients carrying no Bw4 motif on HLA-B which was broader than that of patients carrying 1-2 Bw4 motif(s) on HLA-B [1(0-4)] (P=0.080).(3) The viral loads of HIV-1 infected patients carrying no Bw4 motif on HLA-B were negatively correlated with the magnitude of P24-specific T cell responses (rs=-0.482, P=0.043) and the breadth of responses to P24 peptides (rs=-0.496, P=0.036).No correlations were observed between viral loads and the magnitude or breadth of P24-specific T cell responses in HIV-1 infected patients carrying 1-2 Bw4 motif(s) on HLA-B.Conclusion Compared with HIV-1 infected patients carrying no Bw4 motif on HLA-B, the patients carrying 1-2 Bw4 motif( s) on HLA-B showed lower levels of set point viral load, weak-ened magnitude of P24-specific T cell responses and narrowed breadth of responses to P24 peptides.

Establishment of an enzyme-linked immunosorbent assay for detecting of fumigate hydratase antibody and evaluation of its value in the diagnosis of autoimmune hepatitis / 中华风湿病学杂志

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for detecting human serum fumarate hydratase (FH) antibody and evaluate its role in the diagnosis of autoantigen in autoimmune hepatitis (AIH).Methods The indirect ELISA was established using FH protein,and the reaction conditions were determined.Then,the anti-FH antibody were detected in the serum of 88 AIH patients,56 primary biliary cirrhosis (PBC) patients,50 chronic hepatitis B (HBV) patients,36 chronic hepatitis C (HCV) patients and 98 healthy controls(HC).The results were analyzed with chi-quare and Kruskal-Wallis H methods.Results The ELISA for detecting human anti-FH antibody was established successfully and the optimal reaction conditions were defined.The positive rate of anti-FH antibody in the AIH group (40％) was significantly higher than HC (3％,x2=38.44,P＜0.01),PBC group (7％,x2=18.45,P＜0.01),CHB group (2％,x2=23.59,P＜0.01) and CHC group (6％,x2=14.29,P＜0.01).Anti-FH antibody which was used to diagnose AIH revealed a sensitivity of 40％ and specificity of 94％.Conclusion We have established the ELISA,which is used to detect human anti-FH antibody.It can be detected predominantly in AIH,and this implies that anti-FH antibody may be useful in improving the diagnosis of AIH.

HLA class I alleles and haplotypes in Chinese patients with primary biliary cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To analyze the HLA class I alleles and haplotypes in Chinese patients with primary biliary cirrhosis (PBC).</p><p><b>METHODS</b>Sequencing based typing-polymerase chain reaction (SBT-PCR) was used to investigate the HLA class I alleles of 146 PBC patients and 500 normal controls in northern China. The frequencies of alleles and haplotypes were calculated and compared for the two groups. The chi-square test and Fisher's exact test were used for statistical analyses.</p><p><b>RESULTS</b>There were 26, 51 and 21 alleles identified at the HLA-A, B and C loci respectively, and the frequencies of these alleles were not significantly different between the PBC and normal control groups.However, the frequencies of A *11:01-B*40:06 and A*02:01-B*l5:01 haplotypes were significantly higher in the PBC group than in the normal control group (7.53% vs. 1.40%, P<0.01, OR=5.38; 6.85% vs. 2.00%, P=0.003, OR=3.425).</p><p><b>CONCLUSION</b>This study established the role of HLA class I haplotypes in determining PBC susceptibility in a Chinese population.</p>

Impact of human leukocyte antigen-B specific Bw4 epitope on hepatitis C virus infection / 中华传染病杂志

Objective To study the impact of human leukocyte antigen (HLA)-B specific Bw4 epitope on disease progression of hepatitis C virus (HCV) infection.Methods Eighty-six cases of HCV infection through paid blood donation were enrolled in the study.Enzyme-linked immunosorbent assay (ELISA) to detect HCV IgG and IgM,real-time reverse transcriptation polymerase chain reaction (RT-PCR) to detect HCV RNA,and sequence specific primer-polymerase chain reaction (SSP-PCR) to analyze HLA type was performed.Categorical data were analyzed by chi-square test,and measurement data were compared by independent sample t test.Results Among the 86 HCVinfected individuals,there were 29 (33.7 ％) cases of Bw4/4 homozygote,38 cases (44.2 ％) of Bw4/6 heterozygote and 19 (22.1％) cases of Bw6/6 homozygote.The HCV RNA levels in Bw4/4 group,Bw4/6 group and Bw6/6 group were (3.98±0.32),(5.22±0.29),(5.04±0.38) lg IU/mL,respectively.The HCV RNA level in Bw4/4 group was significantly lower than those in the other two groups (t=2.821,P=0.0063 ; t =2.106,P =0.0407,respectively).The spontaneous clearance rates of Bw4/4 group,Bw4/6 group and BW6/6 group were 58.6％,26.3％ and 21.0％,respectively.The spontaneous clearance rate of Bw4/4 group was significantly higher than those of Bw4/6 group and Bw6/6 group (x2 =7.135,P =0.008; x2 =6.583,P =0.010,respectively).HCV infected individuals with homozygous epitopes of Bw4/4 had 4.351 times higher probability of spontaneous viral clearance than that of Bw4/6 heterozygote or Bw6/6 homozygote (OR=4.351,95％CI:1.676-11.294).Conclusions Homozygosity for HLA-Bw4-bearing B alleles is associated with significant lower HCV viral load and higher spontaneous clearance rate.The HLA-Bw4 epitopes have a protective effect against HCV infection.

The influence of human leucocyte antigen-Ⅰ polymorphisms on plasma viral load in human immunodeficiency virus infected male homosexual population in Beijing / 中华传染病杂志

Objective To analyze the influence of the polymorphisms of human leucocyte antigen (HLA)-Ⅰ molecule and the effects on plasma viral load of human immunodeficiency virus (HIV) infected male homosexual population in Beijing.Methods The HLA-A,HLA-B,HLA-C allele were typed by sequence specific primer-polymerase chain reaction (SSP-PCR),and viral load was detected in 157 chronic HIV infected persons.Normally distributed measurement data were analyzed by one-way or multi-way analysis of variance,while data of abnormal distributions were analyzed by Mann-Whitney U test.Results Among 157 chronic HIV infected persons,the number of Bw4 motifs on HLA-B loci was associated with a lower level of viral load (F=3.01,P=0.045).In these HIV infected persons,the viral load in HLA-B carrying Bw4/4 homozygote was (4.19±0.76) lg IU/mL,in HLA-B carrying Bw6/6 homozygote was (4.63±0.74) lg IU/mL (t=2.27,P=0.010).The viral load of those who carried three,one or none Bw4 motifs on HLA-A and HLA-B loci were (3.92± 0.97),(4.54±0.88) and (4.60±0.72) lg IU/mL,respectively (three vs none:t=2.53,P=0.015; three vs one:t=2.11,P=0.039).HIV infected persons who carried homozygote on any loci of HLA-A,HLA-B,HLA-C had comparable levels of plasma viral load to those who carried heterozygote on HLA-A,HLA-B,HLA-C loci.Among the persons who carried heterozygote on HLA-A,HLA B,HLA-C loci,Bw4/4 homozygote on HLA-B had lower levels of viral load than Bw6/6 homozygote on HLA-B (median:4.09 lg IU/mL vs 4.55 lg IU/mL,U=210.50,P=0.041).HIV infected persons who carried A30/B13/C06 or A33/B58/C03 haplotype had comparable levels of plasma viral load to those without A30/B13/C06 or A33/B58/C03 haplotype (t=0.40,P=0.69; t=0.68,P=0.49,respectively).Conclusions Bw4/4 homozygote on HLA-B loci is associated with lower HIV viral load.Furthermore,the plasma viral load of HIV infected persons carrying heterozygote on HLA-A,HLA-B,HLA-C loci could be influenced by the Bw4/4 homozygote on HLA-B locus,with a lower viral load.

Analysis of influence of HLA-B broad specific Bw4 motif on HCV specific T cell response / 中华微生物学和免疫学杂志

Objective To study the influence of Bw4 broad specific motif on hepatitis C virus (HCV) specific T cell response.Methods The 86 patients with HCV infection were enrolled in this study,who had history of non-standard paid blood donation.The sequence specific primer SSP-PCR and specific primer Amplification Refractory Mutation system was used to analyze HLA type.The T cell response,using PBMCs stimulated by HCV nonstructural protein NS3,NS4 and NS5 was tested by ELISPOT assay.Results There were 29 (33.7％) cases with homozygosity Bw4/4,38 cases (44.2％) with heterozygosity Bw4/6 and 19(22.1％) cases with homozygosity Bw6/6 in 86 patients with HCV infection.HCV viral loads in Bw4/4group,Bw4/6 group and Bw6/6 group were (3.98±0.32) Log (copy/ml),(5.22± 0.29) Log (copy/ml),(5.04±0.38) Log(copy/ml),respectively and there was a significant difference in HCV load among three groups(P=0.0153).The 24 cases with HCV infection were test HCV specific T cell response divided homozygosity Bw4/4 group and non-homozygosity Bw4/4 group.The HCV specific T cell response frequency in homozygosity Bw4/4 group and non-homozygosity Bw4/4 group was 50％ (5/10) and 14.28％ (2/14,respectively.The HCV specific T cell response magnitude in homozygosity Bw4/4 group and non-homozygosity Bw4/4 group was 70 SFU/106 PBMC (0-2020 SFU/106 PBMC)and 0 SFU/106 PBMC (0-200 SFU/106 PBMC).The response magnitude and frequency in homozygosity Bw4/4 group was significantly higher than that of non-homozygosity Bw4/4 group,P value was 0.0450 and 0.069,respectively.In homozygosity Bw4/4 group NS5 induced T cell response was dominant.The NS5 specific T cell response frequency in Bw4/4 group and non-Bw4/4 group was 50％ and 7.14％,respectively,and the difference was nearly significant(P=0.050).Conclusion The HCV-infected blood donors with homozygosity for HLA-Bw4 alleles is associated with a significant lower HCV virus load and stronger HCV specific T cell response,compared with non-homozygosity Bw4/4 group.

The dynamic expression of CD57 on T cell and clinical significance analysis in acute HIV infection / 中华微生物学和免疫学杂志

ObjectiveTo study the dynamic express of CD57 on T cell of PBMC and clinical significance in acute HIV infection.MethodsSeventeen patients with acute HIV infection were enrolled study randomly diagnosed from 2006.11 to 2009.12 and 15 healthy donors as control group.The PBMCs from 1th,3th and 6th during acute infection were collected.The proportion of CD3+CD57+T lymphocytes,CD3+ CD8+CD57 +T lymphocytes and CD3 + CD4 + CD57 + T lymphocytes were evaluated by flow cytometric analysis with three or double color staining.The relationship between the proportion of CD57+ T phenotypes and virus load and CD4+T cells count was analyzed.ResultsThe proportion of CD57+T lymphocytes in PMBC in 1th,3th and 6th during acute HIV acute was 15.24％ ±1.49％,13.51％ ±2.45％ and 14.65％ ±1.83％,respectively,and was higher than normal control group and the difference was significantly(P＜0.0001 ).The proportion of CD8+ CD57+T lymphocytes was 7.79％ ±2.10％ and 9.88％ ±2.36％ in 1th and 3th month during acute infection,respectively.The proportion of CD8+ CD57+T lymphocytes in 1th and 3th month during acute infection were positive relationship with virus load in corresponding time,and R2 was 0.3700 and 0.3768,and P value was 0.0096 and 0.0088,respectively.The proportion of CD8+CD57+T lymphocytes in the 1th and 3th month during acute infection was negative relationship with CD4+T lymphocytes count.The R2 was 0.3768 and 0.4235,and P value was 0.0215 and 0.0017,respectively.In 6 rapid progressors and 11 no rapid progressors on the 1th month after HIV infection,CD8+ CD57+T lymphocytes percentage was 11.20％±2.21％ and 6.16％ ±1.09％,respectively,and CD4+CD57+T lymphocytes percentage 2.79％ ±0.31％and 1.40％ ±0.30％,respectively.Both CD8+CD57+T and CD4+CD57+T in rapid progressors were higher than no rapid progressors,and P value was 0.0338 and 0.0106,respectively.ConclusionCD57+ T lymphocytes percentage in peripheral blood increase in acute HIV infection patients,in which the increasing CD8+CD57+T lymphocyte may mirror the dynamic of HIV replication and CD4+T cell count.The CD57 high express on T lymphocyte on the early HIV acute infection predicts rapid progression.