Objective:To observe the effects of immunosuppressants triptolide(TL)and cyclosporineA(CSA)on HLA antigens expression induced by interferon-γ（INF-γ）in vitro.Method:By using an indirect immunofluorescent method and...Objective:To observe the effects of immunosuppressants triptolide(TL)and cyclosporineA(CSA)on HLA antigens expression induced by interferon-γ（INF-γ）in vitro.Method:By using an indirect immunofluorescent method and analysing with ACAS-570,the abnormal HLA antigen expression of cultured corneal epithelial cells was induced by INF-γ，After incubation with one of the immunosuppressants(CSA,TL) for72hrs.the amount of HLA-ABCand HLA-DRantigens was measured.Result:There was no significant difference(P>0.05)between the group with CAS and the positive control group without CAS.In contrast to CSA,TL dramatically inhibited INF-γinduced expression of HLA antigens of corneal epithelial cells(P<0.001),compared with the control group withoutTL.Conclusion:TL had direct inhibition on the expression of HLA-ABCand HLA-DRantige ns induced by INF-γin vitro,while CSAhad no obvious inhibition,Eye Science2000;16:34-37.展开更多
Dear Editor： Anti-sperm antibodies （ASAs） are composed of numerous antibodies interacting with multiple sperm antigens that play a role in fertility. In males, ASAs cause ＇immune infertility＇ by decreasing sperm ...Dear Editor： Anti-sperm antibodies （ASAs） are composed of numerous antibodies interacting with multiple sperm antigens that play a role in fertility. In males, ASAs cause ＇immune infertility＇ by decreasing sperm counts and normal forms, as well as reducing sperm motility and viability, markedly reducing the likelihood of natural conception. The development of ASA in the male depends on the release of sequestered antigens on germ cells following the disruption of the blood-testis barrier.展开更多
The proliferative response of T-cells to autolo-gous non-T-cells is referred to as the autologous mixed lymphocyte reaction (AMLR). Recent studies have suggested that AMLR represents a mechanism of immune regulation i...The proliferative response of T-cells to autolo-gous non-T-cells is referred to as the autologous mixed lymphocyte reaction (AMLR). Recent studies have suggested that AMLR represents a mechanism of immune regulation in vivo. We investigated AMLR in patients with acute- and chronic myeloid leukemia (AML and CML). AMLR was found to be significantly depressed (P<0.001) in AML patients (n=17, cpm=532±95) and CML patients (n=13, cpm=688±99) when compared with that of their healthy HLA-identical siblings serving as controls (n=17, cpm=4152±619 and n=13 cpm=4086±421, respectively). In order to understand the cellular basis of the defective AMLR in patients with AML end CML, we performed mitogen-treated T-cell cultures analysis of T-cell subsets and HLA-Ⅱ antigen detection on monocytes. The results indicated that the defect of AMLR in patients resided at the stimulator monocyte level rather than at the responder T-cell level. Enumeration of monocytes reactive with monoclonal antibody Tu22, which recognizes determinants of HLA-DQ, demonstrated that ML patients had a significantly decreased (P<0.091) number of circulating Tu22+ monocytes when compared with normal controls. These studies suggest that a deficiency of HLA-DQ+ monocytes contributes to the depression of AMLR in ML and possibly underlies the abnormalities of immune response present in this disease.展开更多
Thirty patients with Vogt-Koyanagi-Haradas disease were typed for HLA-A and HLA-B antigenic determinants by a microlymphocytotoxicity technique. HLA-B22 antigen showed an increased frequency of 43.3% in the patient gr...Thirty patients with Vogt-Koyanagi-Haradas disease were typed for HLA-A and HLA-B antigenic determinants by a microlymphocytotoxicity technique. HLA-B22 antigen showed an increased frequency of 43.3% in the patient group(relative risk=8.69; exact P<0.0001; corrected P<0.0025) compared with normal control group(frequency=7.69%). This association suggests that immunogenetic factor may play an important role in the pathogenesis of Vogt-Koyanagi-Harada's disease.展开更多
In order to evaluate the expression of HLA DR antigen in glandular cells in eutopic and ectopic endometrium in patients with endometriosis, 19 infertile patients with endometriosis were analyzed immunohistoche...In order to evaluate the expression of HLA DR antigen in glandular cells in eutopic and ectopic endometrium in patients with endometriosis, 19 infertile patients with endometriosis were analyzed immunohistochemically by labelled streptavidin biotin (LSAB) method. Nineteen infertile patients without endometriosis were studied as controls. The results showed that the expression of HLA DR antigen in the glandular cells in both eutopic and ectopic endometrium was increased significantly as compared with that in the controls ( P <0.01). It is likely that aberrant expression of HLA DR antigen in endometriotic tissue is involved in abormal immunogenesis of endometriosis.展开更多
In order to study the relationship between replicative status and human leucocyte antigens(HLA),HLA class Ⅱ antigen on hepatocyte membrane was analyzed in liver biopsies from 49 pa-tients with chronic hepatitis B vir...In order to study the relationship between replicative status and human leucocyte antigens(HLA),HLA class Ⅱ antigen on hepatocyte membrane was analyzed in liver biopsies from 49 pa-tients with chronic hepatitis B virus infection.The results revealed that expression of HLA classⅡ antigen on hepatocyte membrane was observed in 57% (13/23) of hepatitis B e antigen (HBeAg)positive,only in 15% (4/26) of anti-HBe positive carriers.The display of HLA class Ⅱ antigen onhepatocyte membrane yeas found in 65% (11/17) hepatitis B core antigen (HBcAg) positive and in19% (6/32) HBcAg negative patients with chronic hepatitis B virus infection.There was nosignificant difference in the expression of HLA class Ⅱ antigen on hepatocyte membrane between mi-nor hepatic diseases and active liver diseases.These findings suggested that display of HLA classⅡ antigen on hepatocyte membrane might be associated with viral replication in the chronic hepati-tis B virus carriers.展开更多
This study was designed to establish the prevalence of HLA-B*5701 at HIV-1 infected individuals in Brazil. A total of 517 consecutive individuals were followed in this study from February 2009 through July 2011. The p...This study was designed to establish the prevalence of HLA-B*5701 at HIV-1 infected individuals in Brazil. A total of 517 consecutive individuals were followed in this study from February 2009 through July 2011. The presence of HLA-B*5701 was determined by Nested-PCR with HLA-B*57 and HLA-B*5701 sequence-specific primers (PCR-SSP). The expression of HLA-B*57 was negative in the 385 (74.5%) and positive in the 103 (19.9%) of infected individuals. Among these, the expression of HLA-B5701 was positive in the 29 (5.6%) of individuals. No demographic or ethnic differences were found between HLA-B*57/HLA-B*5701 HIV-1 negative patients, with a prevalence of Caucasians (57.1%) individuals. During the period of study, 68 patients were submited to an abacavir contain- ing regimen. The HLA-B*5701 allele was observed in 7 (10.3%) patients, with a significant incidence of Hypersensitivity reactions at 4 of them (p < 0.001). Conclusions: Although Brazilian population consists of a mixture of individuals of Caucasian, African and Native American genetic background, prevalence of HLA-B*5701 in this population is similar to the one found in pure Caucasians.展开更多
40 thyroid specimens from Tru-cut needle biopsy and surgerywere analyzed by APAAP immunohistochemical technique.It was shown thatthe expression of HLA-DR antigen on thyroid follicular cells (TFC) wasinvolved in almost...40 thyroid specimens from Tru-cut needle biopsy and surgerywere analyzed by APAAP immunohistochemical technique.It was shown thatthe expression of HLA-DR antigen on thyroid follicular cells (TFC) wasinvolved in almost every patient.The degree of DR expression was signifi-cantly higher in autoimmune thyroiditis (AIT) than in Graves’ disease (GD)and in nontoxic goiter (NTG),The level of DR expression on TFC not onlycorrelated markedly with degree of MNC infiltration,but more so with degreeof DR expression on intrathyroidal infiltrates,suggesting that aberrant DR ex-pression in vivo may be related to the activation of intrathyroidal T cells.展开更多
AIM: To establish a simplified method for generating peptide-major histocompatibility complex (MHC) class I tetramers.METHODS: cDNAs encoding the extracellular domain of human lymphocyte antigen (HLA)-A*0201 heavy cha...AIM: To establish a simplified method for generating peptide-major histocompatibility complex (MHC) class I tetramers.METHODS: cDNAs encoding the extracellular domain of human lymphocyte antigen (HLA)-A*0201 heavy chain (A2) and β2-microglobulin (β2m) from total RNA extracted from leukocytes of HLA-A2+ donors were doned into separate expression vectors by reverse transcription-polymerase chain reaction. The recombinant A2 and β2m proteins were expressed in Escherichia coli strain BL21(DE3) and recovered from the inclusion body fraction. Soluble A2 proteins loaded with specific antigen peptides were refolded by dilution from the heavy chain in the presence of light chain β2m and HLA-A2-restricted peptide antigens. The refolded A2monomers were biotinylated with a commercial biotinylation enzyme (BirA) and purified by low pressure anion exchange chromatography on a Q-Sepharose (fast flow) column.The tetramers were then formed by mixing A2 monomers with streptavidin-PE in a molar ratio of 4:1. Flow cytometry was used to confirm the expected tetramer staining of CD8+ T cells.RESULTS: Recombinant genes for HLA-A*0201 heavy chain (A2) fused to a BirA substrate peptide (A2-BSP) and mature β2m from HLA-A2+ donor leukocytes were successfully doned and highly expressed in E. coli. Two soluble monomeric A2-peptide complexes were reconstituted from A2-BSP in the presence of β2m and peptides loaded with either human cytomegalovirus pp65495-503 peptide (NLVPMVATV,NLV; designated as A2-NLV) or influenza virus matrix protein Mp58-66 peptide (GILGFVFTL, GIL; designated as A2-GIL). Refolded A2-NLV or A2-GIL monomers were biotinylated and highly purified by single step anion exchange column chromatography. The tetramers were then formed by mixing the biotinylated A2-NLV or A2-GIL monomers with streptavidin-PE, leading to more than 80% multiplication as revealed by SDS-PAGE under non-reducing, unboiled conditions. Flow cytometry revealed that these tetramers could specifically bind to CD8+ T cells from a HLA-A2+ donor,but failed to bind to those from a HLA-A2- donor.CONCLUSION: The procedure is simple and efficient for generating peptide-MHC tetramers.展开更多
Objective:To compare the genotype frequencies of HLA class-ⅡDRB1 alleles in Giardia(G.)lamblia-infected children.Methods:A total of 490 Egyptian children aged 2-16 years were subjected to microscopic stool examinatio...Objective:To compare the genotype frequencies of HLA class-ⅡDRB1 alleles in Giardia(G.)lamblia-infected children.Methods:A total of 490 Egyptian children aged 2-16 years were subjected to microscopic stool examination to detect G.lamblia infection,and to exclude other intestinal pathogens.On the basis of their microscopic findings,a group of 80 children were chosen as giardiasis cases,another 80 children were confirmed as Giardia free control group by immunochromatographic test,and the remaining children were excluded.Both giardiasis and control groups were then subjected to blood examination to identify their genetic type of HLA-DRB1 alleles.Results:HLA class-ⅡDRB1*03:01 and DRB1*13:01 alleles were significantly associated with G.lamblia infection(P<0.001 for each variable).On the other hand,HLA class-ⅡDRB1*04:02,DRB1*10:01,DRB1*14:01 and DRB1*15:01 alleles were significantly demonstrated in Giardia free children.However,other HLA-DRB1 alleles did not show any significant association with giardiasis.Conclusions:HLA class-ⅡDRB1*03,DRB1*13,DRB1*04,DRB1*10,DRB1*14 and DRB1*15 alleles may be involved in the establishment of host immune response to G.lamblia infection.展开更多
文摘Objective:To observe the effects of immunosuppressants triptolide(TL)and cyclosporineA(CSA)on HLA antigens expression induced by interferon-γ（INF-γ）in vitro.Method:By using an indirect immunofluorescent method and analysing with ACAS-570,the abnormal HLA antigen expression of cultured corneal epithelial cells was induced by INF-γ，After incubation with one of the immunosuppressants(CSA,TL) for72hrs.the amount of HLA-ABCand HLA-DRantigens was measured.Result:There was no significant difference(P>0.05)between the group with CAS and the positive control group without CAS.In contrast to CSA,TL dramatically inhibited INF-γinduced expression of HLA antigens of corneal epithelial cells(P<0.001),compared with the control group withoutTL.Conclusion:TL had direct inhibition on the expression of HLA-ABCand HLA-DRantige ns induced by INF-γin vitro,while CSAhad no obvious inhibition,Eye Science2000;16:34-37.
文摘Dear Editor： Anti-sperm antibodies （ASAs） are composed of numerous antibodies interacting with multiple sperm antigens that play a role in fertility. In males, ASAs cause ＇immune infertility＇ by decreasing sperm counts and normal forms, as well as reducing sperm motility and viability, markedly reducing the likelihood of natural conception. The development of ASA in the male depends on the release of sequestered antigens on germ cells following the disruption of the blood-testis barrier.
文摘The proliferative response of T-cells to autolo-gous non-T-cells is referred to as the autologous mixed lymphocyte reaction (AMLR). Recent studies have suggested that AMLR represents a mechanism of immune regulation in vivo. We investigated AMLR in patients with acute- and chronic myeloid leukemia (AML and CML). AMLR was found to be significantly depressed (P<0.001) in AML patients (n=17, cpm=532±95) and CML patients (n=13, cpm=688±99) when compared with that of their healthy HLA-identical siblings serving as controls (n=17, cpm=4152±619 and n=13 cpm=4086±421, respectively). In order to understand the cellular basis of the defective AMLR in patients with AML end CML, we performed mitogen-treated T-cell cultures analysis of T-cell subsets and HLA-Ⅱ antigen detection on monocytes. The results indicated that the defect of AMLR in patients resided at the stimulator monocyte level rather than at the responder T-cell level. Enumeration of monocytes reactive with monoclonal antibody Tu22, which recognizes determinants of HLA-DQ, demonstrated that ML patients had a significantly decreased (P<0.091) number of circulating Tu22+ monocytes when compared with normal controls. These studies suggest that a deficiency of HLA-DQ+ monocytes contributes to the depression of AMLR in ML and possibly underlies the abnormalities of immune response present in this disease.
文摘Thirty patients with Vogt-Koyanagi-Haradas disease were typed for HLA-A and HLA-B antigenic determinants by a microlymphocytotoxicity technique. HLA-B22 antigen showed an increased frequency of 43.3% in the patient group(relative risk=8.69; exact P<0.0001; corrected P<0.0025) compared with normal control group(frequency=7.69%). This association suggests that immunogenetic factor may play an important role in the pathogenesis of Vogt-Koyanagi-Harada's disease.
文摘In order to evaluate the expression of HLA DR antigen in glandular cells in eutopic and ectopic endometrium in patients with endometriosis, 19 infertile patients with endometriosis were analyzed immunohistochemically by labelled streptavidin biotin (LSAB) method. Nineteen infertile patients without endometriosis were studied as controls. The results showed that the expression of HLA DR antigen in the glandular cells in both eutopic and ectopic endometrium was increased significantly as compared with that in the controls ( P <0.01). It is likely that aberrant expression of HLA DR antigen in endometriotic tissue is involved in abormal immunogenesis of endometriosis.
文摘In order to study the relationship between replicative status and human leucocyte antigens(HLA),HLA class Ⅱ antigen on hepatocyte membrane was analyzed in liver biopsies from 49 pa-tients with chronic hepatitis B virus infection.The results revealed that expression of HLA classⅡ antigen on hepatocyte membrane was observed in 57% (13/23) of hepatitis B e antigen (HBeAg)positive,only in 15% (4/26) of anti-HBe positive carriers.The display of HLA class Ⅱ antigen onhepatocyte membrane yeas found in 65% (11/17) hepatitis B core antigen (HBcAg) positive and in19% (6/32) HBcAg negative patients with chronic hepatitis B virus infection.There was nosignificant difference in the expression of HLA class Ⅱ antigen on hepatocyte membrane between mi-nor hepatic diseases and active liver diseases.These findings suggested that display of HLA classⅡ antigen on hepatocyte membrane might be associated with viral replication in the chronic hepati-tis B virus carriers.
文摘This study was designed to establish the prevalence of HLA-B*5701 at HIV-1 infected individuals in Brazil. A total of 517 consecutive individuals were followed in this study from February 2009 through July 2011. The presence of HLA-B*5701 was determined by Nested-PCR with HLA-B*57 and HLA-B*5701 sequence-specific primers (PCR-SSP). The expression of HLA-B*57 was negative in the 385 (74.5%) and positive in the 103 (19.9%) of infected individuals. Among these, the expression of HLA-B5701 was positive in the 29 (5.6%) of individuals. No demographic or ethnic differences were found between HLA-B*57/HLA-B*5701 HIV-1 negative patients, with a prevalence of Caucasians (57.1%) individuals. During the period of study, 68 patients were submited to an abacavir contain- ing regimen. The HLA-B*5701 allele was observed in 7 (10.3%) patients, with a significant incidence of Hypersensitivity reactions at 4 of them (p < 0.001). Conclusions: Although Brazilian population consists of a mixture of individuals of Caucasian, African and Native American genetic background, prevalence of HLA-B*5701 in this population is similar to the one found in pure Caucasians.
文摘40 thyroid specimens from Tru-cut needle biopsy and surgerywere analyzed by APAAP immunohistochemical technique.It was shown thatthe expression of HLA-DR antigen on thyroid follicular cells (TFC) wasinvolved in almost every patient.The degree of DR expression was signifi-cantly higher in autoimmune thyroiditis (AIT) than in Graves’ disease (GD)and in nontoxic goiter (NTG),The level of DR expression on TFC not onlycorrelated markedly with degree of MNC infiltration,but more so with degreeof DR expression on intrathyroidal infiltrates,suggesting that aberrant DR ex-pression in vivo may be related to the activation of intrathyroidal T cells.
基金Supported by the National Natural Science Foundation of China, No. 30230350 and No. 30371651Major State Basic Research Development Program of China, 973 Program, No. G2000057006
文摘AIM: To establish a simplified method for generating peptide-major histocompatibility complex (MHC) class I tetramers.METHODS: cDNAs encoding the extracellular domain of human lymphocyte antigen (HLA)-A*0201 heavy chain (A2) and β2-microglobulin (β2m) from total RNA extracted from leukocytes of HLA-A2+ donors were doned into separate expression vectors by reverse transcription-polymerase chain reaction. The recombinant A2 and β2m proteins were expressed in Escherichia coli strain BL21(DE3) and recovered from the inclusion body fraction. Soluble A2 proteins loaded with specific antigen peptides were refolded by dilution from the heavy chain in the presence of light chain β2m and HLA-A2-restricted peptide antigens. The refolded A2monomers were biotinylated with a commercial biotinylation enzyme (BirA) and purified by low pressure anion exchange chromatography on a Q-Sepharose (fast flow) column.The tetramers were then formed by mixing A2 monomers with streptavidin-PE in a molar ratio of 4:1. Flow cytometry was used to confirm the expected tetramer staining of CD8+ T cells.RESULTS: Recombinant genes for HLA-A*0201 heavy chain (A2) fused to a BirA substrate peptide (A2-BSP) and mature β2m from HLA-A2+ donor leukocytes were successfully doned and highly expressed in E. coli. Two soluble monomeric A2-peptide complexes were reconstituted from A2-BSP in the presence of β2m and peptides loaded with either human cytomegalovirus pp65495-503 peptide (NLVPMVATV,NLV; designated as A2-NLV) or influenza virus matrix protein Mp58-66 peptide (GILGFVFTL, GIL; designated as A2-GIL). Refolded A2-NLV or A2-GIL monomers were biotinylated and highly purified by single step anion exchange column chromatography. The tetramers were then formed by mixing the biotinylated A2-NLV or A2-GIL monomers with streptavidin-PE, leading to more than 80% multiplication as revealed by SDS-PAGE under non-reducing, unboiled conditions. Flow cytometry revealed that these tetramers could specifically bind to CD8+ T cells from a HLA-A2+ donor,but failed to bind to those from a HLA-A2- donor.CONCLUSION: The procedure is simple and efficient for generating peptide-MHC tetramers.
文摘Objective:To compare the genotype frequencies of HLA class-ⅡDRB1 alleles in Giardia(G.)lamblia-infected children.Methods:A total of 490 Egyptian children aged 2-16 years were subjected to microscopic stool examination to detect G.lamblia infection,and to exclude other intestinal pathogens.On the basis of their microscopic findings,a group of 80 children were chosen as giardiasis cases,another 80 children were confirmed as Giardia free control group by immunochromatographic test,and the remaining children were excluded.Both giardiasis and control groups were then subjected to blood examination to identify their genetic type of HLA-DRB1 alleles.Results:HLA class-ⅡDRB1*03:01 and DRB1*13:01 alleles were significantly associated with G.lamblia infection(P<0.001 for each variable).On the other hand,HLA class-ⅡDRB1*04:02,DRB1*10:01,DRB1*14:01 and DRB1*15:01 alleles were significantly demonstrated in Giardia free children.However,other HLA-DRB1 alleles did not show any significant association with giardiasis.Conclusions:HLA class-ⅡDRB1*03,DRB1*13,DRB1*04,DRB1*10,DRB1*14 and DRB1*15 alleles may be involved in the establishment of host immune response to G.lamblia infection.
