Kinetics of the pregnancy-induced humoral and cellular immune response against the paternal HLA class I antigens of the child

Pregnancy can prime the maternal humoral immune response against paternal human leukocyte antigens (HLA) of the child. Previous studies have reported that formation of antibodies against inherited paternal HLA is associated with the presence of primed cytotoxic T lymphocytes (CTLs) specific for these antigens. Recently, we reported that primed CTLs can persist for more than 10 years after pregnancy even if the antibodies have disappeared. In the present study we studied the kinetics of the pregnancy induced immune response of the T-cell and B-cell compartment. In 12 women, who had specific antibodies against the paternal HLA antigens of the child (child mismatch) at the time of delivery, we analyzed the CTLp frequencies against the paternal HLA antigens from the time of delivery up to 2 years after. The contribution of primed CTLs to these CTLp frequencies was tested by limiting dilution analysis in the absence and presence of monoclonal antibodies specific for CD8. In contrast to na?ve CTLs, primed CTLs are resistant to CD8 antibodies. Disappearance of the antibodies was not associated with a decrease of the number of CTLp directed against the paternal antigens, towards which the antibodies were originally directed. However, in women where the antibodies disappeared, a decrease of primed child mismatch specific CTLs was found, whereas in women where the antibodies persist, the population of primed CTLs remained stable up to 2 years after delivery. Our data suggest a functional correlation between the T-cell and B-cell allorepertoire. Although the kinetics do not run completely in parallel, disappearance of the anti-HLA antibodies in the first 2 years after delivery is related with a decrease of primed child mismatch specific CTLs. These data may be relevant for transplantation of female recipients with historical, pregnancy-induced HLA alloantibodies.     

胃癌组织HLA－I类和DR分子免疫组化研究

本文应用免疫组化法对６４例胃癌、癌旁组织和６例胃溃疡大致正常胃粘膜冰冻和石蜡切片进行了染色．结果表明，正常胃粘膜和癌旁胃粘膜上皮细胞ＨＬＡ－Ｉ类分子表达阳性，其着色较均一，ＨＬＡ－ＤＲ染色均阴性．胃癌细胞Ｉ类分子表达缺失（２７／６４例），与癌旁上皮比较差异显著（Ｐ＜０．０１）。粘液细胞癌和低分化癌Ｉ类分子缺失率显著高于高分化癌（Ｐ＜０．０２５）．此外，发生肿瘤转移的病例Ｉ类分子缺失率（１２／１５例）显著高于无转移组（１／５例，Ｐ＜０．０２５）．ＤＲ分子在癌组织表达阳性，其阳性率高达５３．１％（３４／６４例）．低分化癌ＤＲ分子阳性率亦显著高于高分化癌和中分化癌，未分化癌ＤＲ分子阳性率亦显著高于高分化癌（Ｐ＜０．０１～０．０５）．提示（１）ＨＬＡ－Ｉ类分子表达缺失可能与癌细胞逃避宿主免疫监视发生润浸生长和转移有关；（２）分化程度不同的癌组织ＨＬＡ－Ｉ类分子表达差异显著，提示癌细胞分化可能影响Ｉ、Ⅱ类分子表达和肿癌抗原呈递；（３）ＨＬＡ－Ｉ类和ＤＲ分子表达异常可能是上皮恶性转变的标志之一．     

HLA-DR antigens and the antibody response against Epstein-Barr virus

Antibodies against Epstein-Barr virus (EBV) antigens, i.e. the viral capsid antigen (VCA) and the Epstein-Barr nuclear antigen (EBNA), were determined in two independent populations with known HLA-DR phenotypes. The first population consisted of 151 patients with rheumatoid arthritis; the second one of 88 healthy parents of leukemic children. Although the group of patients with rheumatoid arthritis differed significantly in the frequency of 4 DR antigens from the second group, both groups had the same correlation between HLA-DR antigens and the antibody response to EBV antigens. There was a significant correlation between HLA-DR1 and reduced titers of antibodies to VCA, whereas the persons with only one identifiable DR antigen had higher anti-VCA titers. The persons with HLA-DR5 had significantly higher anti-EBNA titers than those without DR5.     

HLA class I antigens in severe RSV bronchiolitis

Cytotoxic T-lymphocytes (CTL) recognize virus-infected cells in association with HLA class I antigens. There is strong evidence of the importance of CTL in respiratory syncytial virus (RSV) infections. We looked for but were unable to demonstrate an association between particular HLA class I antigens and severe RSV bronchiolitis in infants.     

Aging: the cellular immune response against autologous and foreign antigens is affected before the humoral response

In this study we examined the auto- and hetero-immune response in mice of different ages immunized with antigens of Trypanosoma cruzi (S-105). We observed that 20-day- and 12-month-old mice showed decreased response to foreign antigens and increased response to autoantigens, compared with 3-month-old immunized mice. The 6-month-old mice showed hetero- and autoimmune cellular responses similar to those of 12-month-old animals; however, the humoral response was similar to that of 3-month-old animals against either antigen, suggesting that the compartments of the immune response are altered at different moments in the same individual. Immune response against a foreign antigen is correlated with the presence of cellular infiltrate in skeletal and heart muscle whereas no modifications in the tissue are noticed in animals with an autoimmune response. Also, we observed from cell transfer experiments that lymph node cells are involved in the dysregulation that we noticed with aging.     

Differential immunogenicity of HLA class I alloantigens for the humoral versus the cellular immune response: "towards tailor-made HLA mismatching"

The immunogenicity of an individual human leukocyte antigen (HLA) class I mismatch is different for the cellular and the humoral alloimmune responses. The consequence is that the same antigen can induce a strong antibody response and no cytotoxic T lymphocyte reactivity, but the reverse can occur also. Exact knowledge of the immunogenicity of an HLA mismatch for an individual patient can lead to a strategy of tailor-made HLA mismatching if no HLA identical donor is available. Depending on the clinical situation, one should select a donor with HLA mismatches according to the humoral or cellular mismatch algorithm.     

The expression of HLA class I antigens in germ cell testicular cancer

The expression of HLA class I antigens in testicular germ cell tumors (TGCTs) was studied by the immunoperoxidase technique. In the normal testicle, the interstitial cells of Leydig as well as most of the germ cells were significantly stained. In typical seminoma, 75% of the tumor cells in stage I and 30% in stage II were stained. In embryonal cell carcinoma, 25% of the cases in stage I and less than 10% of those in stage II were stained. Mature teratoma was stained in most of the cases, whereas in malignant teratoma only 35% of the cases showed some staining of the tumor cells. In mixed tumors each component displayed its characteristic staining pattern. The expression of class I antigens on tumor cells is required for immune recognition and lysis of the tumor by cytotoxic T-cells. The reduced expression of class I antigens that was related to histologic characteristics and stage suggests that some testicular tumors may escape immune surveillance and become biologically more aggressive.     

Comparison of the humoral and cytotoxic T-lymphocyte response to individual HLA class I alloantigens in highly immunized patients

In order to investigate the correlation between activation of cytotoxic T-lymphocyte precursor (CTLp) and the formation of antibodies to alloantigens, we studied 21 highly sensitized patients waiting for a kidney transplantation. Both antibody reactivity and CTLp frequencies of these patients were determined against 88 individual HLA class I alloantigens. A high or low CTLp frequency against a certain HLA-A or -B alloantigen was not correlated with the presence or absence of the antibodies to that antigen. Mismatched antigens, towards which the patient had not formed antibodies, can induce either a higher or a lower frequency of CTLp compared to mismatches towards which the patients had formed antibodies. The possible implications of this lack of correlation between the T- and B-cell allorepertoires with regard to donor selection for (highly immunized) patients is discussed.     

Soluble HLA-DR molecules in patients with HLA class II versus class I associated disorders

HLA genes have been identified as key genetic factors contributing to many chronic diseases characterized by autoimmune features. The role of HLA encoded molecules in the pathogenesis of these diseases is unresolved. We have now analysed soluble HLA-DR molecules circulating in the serum of patients with different autoimmune diseases and have defined parameters controlling serum levels. Patients with HLA-DR associated diseases were characterized by elevated serum concentrations of HLA-DR molecules and were clearly distinct from patients with HLA-B27 associated disorders. We did not find evidence for a correlation between disease activity, laboratory abnormalities and elevated serum concentrations of soluble HLA-DR molecules. Studies in normal individuals indicated that soluble HLA-DR molecules are at least partially regulated by the HLA haplotype. Highest serum concentrations were found in individuals carrying the HLA-DR3 or HLA-DR4 haplotype raising the possibility that the phenomenon of HLA-disease association reflects differences in the genetic control of soluble HLA-DR molecules. Interferon-gamma treatment caused an increase in serum concentrations of soluble HLA-DR molecules, whereas a decrease of circulating HLA-DR molecules was associated with an immunosuppressive with cyclosporine A. These data suggest that the patient's immunoresponsiveness represents a second important mechanism controlling circulating HLA-DR molecules.     

Obtaining a highly purified preparation of class I HLA antigens

Institute of Molecular Biology, Academy of Sciences of the USSR. Institute of Carcinogenesis, All-Union Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. N. Trapeznikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 110, No. 10, pp. 401–402, October, 1990.     

The goldfish immune response. I. Characterization of the humoral response to particulate antigens.

Anti-red blood cells (RBC) and anti-hapten antibody synthesis were studied in the goldfish, Carassius auratus. Spontaneous haemagglutination titres were found against all the antigens tested. A weak secondary response was observed in RBC-primed fish boosted during the end-phase of the primary antibody production. However, when the second antigenic challenge was performed during the early exponential phase of a primary stimulation, an important amplified response was obtained. The antibody production and immunological memory can be dissociated: no antibody synthesis occurred in glutaraldehyde-fixed RBC (F-RBC) primed was obtained when untreated or F-RBC were given to F-RBC primed animals. The amplified response to sheep red blood cells (SRBC) was significantly inhibited when fish were primed with a mixture of SRBC and Xenopus red blood cells (XRBC), demonstrating an antigenic competition phenomenon. Studies on anti-trinitrobenzene responses confirm the efficiency of E. coli lipopolysaccharide as a carrier for fish anti-hapten immunization. The kinetics and regulation of antibody synthesis in fish are discussed in relation to the described results.     

Correlations between Terasaki's HLA class II epitopes and HLAMatchmaker-defined eplets on HLA-DR and -DQ antigens

Human leukocyte antigen (HLA) class II-specific antibodies increase the risk of transplant failure, and their characterization must consider epitopes rather than antigens. There are two strategies to determine HLA epitope structure. Terasaki's group has analyzed antibody reactivity patterns with single antigen panels with a computer program based on shared amino acid residues of reactive alleles. HLAMatchmaker is a theoretical algorithm that predicts HLA epitopes on the HLA molecular surface from stereochemical modeling of epitope–paratope interfaces of antigen–antibody complexes. Our epitope repertoire is based on so-called 'eplets' representing 3-Å patches of at least one polymorphic residue on the molecular surface. This report describes how 49 of 53 Terasaki's HLA-DR epitopes correspond to HLAMatchmaker-defined eplets. Most of them are equivalent to single eplets ( n  = 33) or two or more possible eplets ( n  = 10), but six had corresponding eplet pairs. There were 10 cases whereby eplets have permissible residue combinations, and in 5 cases, we found that eplet specificity might be influenced by nearby hidden residues. We could assign corresponding eplets to 17 of 18 Terasaki's HLA-DQ epitopes. This study demonstrates how the HLAMatchmaker interpretation of amino acid residues shared between antibody-reactive antigens can increase our understanding of the structural basis of HLA epitopes.     

DNA typing for class II HLA antigens with allele-specific or group-specific amplification. I. Typing for subsets of HLA-DR4

DNA sequences that distinguish the subsets of HLA-DR4 are also found on several other alleles. This makes typing of heterozygotes with oligonucleotide probes quite impractical. We have therefore developed a procedure in which, in a first step, DNA of the genes to be analyzed is amplified selectively, using group-specific primers. In the case of DR4-DRB1, a primer matching codons 5 to 13 when used in the polymerase chain reaction resulted in products that were entirely suitable for typing for the DR4 subsets. Using appropriate probes, eight distinct subsets were identified. However, only six of them were represented in a normal North American Caucasian panel. In conjunction with a method for rapid DNA extraction, the procedure offers a simple, highly specific and reproducible method for determining subtypes of HLA-DR4 that at present cannot be recognized by serologic methods.     

Soluble HLA class I and class II antigens in patients with multiple sclerosis

Abstract: Soluble HLA class I (sHLA-I) and soluble HLA class II (sHLA-II) antigen levels during different stages of disease were investigated in paired serum and cerebrospinal fluid (CSF) samples from 37 patients with multiple sclerosis (MS) using ELISA and Western blot analysis. Soluble HLA-II antigens in the serum of untreated patients with the relapsing-remitting type of MS (RRMS) were found to be significantly elevated in acute relapse as compared to values obtained from patients under steroid treatment, in remission or healthy controls. No significant differences in circulating sHLA-I levels could be detected. In contrast, a trend towards increased intrathecal production of sHLA-I molecules in the CSF was observed in untreated RRMS patients in acute relapse, whereas the levels of soluble HLA-II antigens in the CSF were below the detection limit of the ELISA method. Our observations underline the presence of systemic immune activation in MS patients, as reflected in elevated serum sHLA-II antigen levels, and reveal a dichotomy between sHLA class I and II antigen production in the peripheral blood versus CSF in acute MS. Serial measurements of sHLA-II antigen levels might represent a non-invasive method to assess disease activity in MS patients.     

Expression of HLA class I antigens on hepatocytes in liver disease

Recent studies have demonstrated aberrant expression and topographical heterogeneity of HLA Class I and Class II antigens in tissues of patients with certain immunologic or neoplastic diseases. Current information about the expression of HLA antigens by normal and diseased hepatocytes is controversial. We analyzed the HLA antigenic profile of 4 normal fetal livers, 5 normal adult livers, 7 cases of chronic hepatitis B virus (HBV) infection, 14 cases of cirrhosis of various etiologies, 11 hepatic neoplasms, and 5 continuous cell lines derived from hepatic tumors. The specimens were tested by the indirect fluorescent antibody method with a panel of monoclonal antibodies to distinct monomorphic determinants of HLA Class I and Class II antigens and to beta 2-microglobulin. HLA Cells I antigens were not detected on normal fetal and adult hepatocytes, but were displayed on the plasma membrane of hepatocytes in the majority of all hepatic diseases tested and of the 5 hepatic tumor cell lines. There was a significant correlation between the expression of HLA Class I antigens on hepatocytes and the intensity of intralobular inflammation. Double immunofluorescent staining of livers infected with hepatitis B virus demonstrated simultaneous expression of HLA Class I antigens and HBsAg or HBcAg only in a small percentage of hepatocytes, suggesting lack of a specific association between HLA Class I and these viral antigens. HLA Class II antigens were not detected on hepatocytes from any of the liver diseases tested but were expressed by one of the 5 liver carcinoma cell lines analyzed. These findings confirm that HLA Class I antigens are not detectable within the limits of several immunohistochemical methods on normal hepatocytes and suggest that injury by a variety of factors directly or indirectly leads to induction of these antigens on the plasma membrane of hepatocytes.     

Soluble HLA class I and HLA-DR plasma levels in patients with anterior uveitis

Anterior uveitis (AU) is an autoimmune disease frequently associated with HLA-B27 antigen. Because of the immune regulatory properties of soluble human leukocyte antigen (sHLA) molecules, we quantified sHLA class I (sHLA-I) and sHLA-DR plasma levels in HLA-typed AU patients (n = 60). Randomly selected healthy individuals (n = 128) and HLA-B27 antigen-positive individuals (n = 24) with HLA phenotype frequencies similar to the HLA-B27 antigen-positive AU patients served as control panels. As expected, HLA-B27 phenotype was significantly increased in AU patients (n = 60), compared to healthy controls. Mean sHLA-I levels in AU patients were slightly higher than in randomly selected healthy controls. Regarding AU subgroups, elevated sHLA-I levels were only found in HLA-B27 antigen-negative patients. Compared to controls, sHLA-DR levels were significantly increased in AU patients and the subgroups of HLA-B27 antigen-negative and -positive patients but not Fuchs' heterochromic cyclitis (FHC). AU patients negative for HLA-B27 antigen with a chronic course had higher sHLA-DR levels than those with an acute course. The presence of associated systemic diseases in AU patients was related to elevated sHLA-DR levels. Secretion of sHLA-DR in blood differs among the various forms of AU. Systemic immune activation was present in AU but not in FHC.     

Both class I and class II HLA antigens are thyroid cancer susceptibility factors

Abstract: It has been established that HLA antigens are susceptibility factors for different cancers, including thyroid tumors. However, the diversity and sometimes weak and contradictory associations found have frequently led to the view that the HLA and tumorigenesis links might be the result of statistical errors. However, it has recently been established that it is indeed a currently complex and unexplained but real phenomenon, which may be crucial in preventing several types of cancer. In the present work we have found in a relatively large series of thyroid cancer patients (n = 161) that both HLA class I (B35) and class II (DR11) antigens are susceptibility factors only in the papillary tumor group of patients, B35 association p value is found at the limit of significance (p_c(120) = 0.05); the follicular group did not show any HLA association, suggesting that the etiopathogenesis of each type of cancer is different. HLA-B35 and DR11 are not working together to induce tumorigenesis and each of them seems to confer susceptibility by using different pathways or by being markers of distinct neighboring susceptibility genes. DR4 has also been found in 86% (n = 6) of Hürthle cell carcinoma. No association has been found between HLA and disease activity. HLA mechanisms of association to cancer are discussed and a world-wide HLA/tumorigenic study is proposed to obtain a clear picture of the puzzling and controversial susceptibility markers found in different tumors and in different ethnic groups.     

Interethnic genetic differentiation: HLA class I antigens in the population of Mongolia

A total of 1668 individuals representing 10 major Mongolian ethnic groups were serologically typed for HLA-A, -B, and -C antigens. Antigens A2, A24, B61, B51, B58, Cw3, Cw7, and Cw6 were the most frequent specificities in Mongolians and no case of B42 was noted in all ethnic groups. The cluster analysis of Principal Components I and II shows that Mongolian speaking groups form one cluster vs Turkic-speaking Kazakhs. The analysis reveals a low, but significant differentiation of Mongolian ethnic groups as measured by F_ST = 0.0100 (P < 0.001). Gene diversity analysis shows that the genetic diversity of the Mongolian population can be attributed largely to its ethnic component, which makes up 64% of total genetic variation. The low degree of interpopulation variation and high level of intrapopulation diversity can be explained by the nomadic way of life of this indigenous population. Three-locus haplotypes A24-B61-Cw3, A33-B58-Cw3 are the most common haplotypic associations in Mongolians. The presence of antigens characteristic of Mongoloid, Caucasoid, and Negroid populations in Mongolians suggests a unique genetic background of this indigenous population. The three-locus haplotype distribution among Mongolians relative to other world populations supports the migration of ancient people from Central Asia to the New World, Korean Peninsula, and Southeast Asia. Am. J. Hum. Biol. 11:603–618, 1999. © 1999 Wiley-Liss, Inc.     

Immune response to H-2 class I antigens on platelets. I. Immunogenicity of platelet class I antigens

Purified allogeneic murine platelet suspensions were found unable to induce primary anti-H-2 class I antibody or T cell proliferative responses. In contrast, the same platelet suspensions could elicit secondary anti-class I responses. The secondary responses were not due to contaminating leucocytes. Possible explanations, the lack of acolyte determinants (class II or non- H-2) on platelets or inappropriate layout and/or structure of their class I antigens, are discussed. These findings emphasize the importance of sufficient leucocyte depletion before platelet transfusion in the human.