Showing papers in "Disease Markers in 1990"

Changes in the expression of HLA-class II antigens on peripheral blood monocytes from aged humans.

Abstract: Increased incidence of infections, cancer, monoclonal gammopathies and rheumatic diseases in aged humans has been described. Histocompatibility antigens are involved in the regulation of immune response and it has been suggested that age-related alterations in the murine immune system may be due to changes in the expression of these antigens on the immunocompetent cells. In this paper we study the expression of HLA-DR/DP and HLA-DQ antigens on monocytes from healthy human elderly donors. Results show that peripheral blood monocytes from elderly subjects express decreased levels of HLA-DR/DP antigens (61.5 + 16.3) when compared to young controls (82.5 + 8.5) and increased levels of HLA-DQ antigens (40.4 + 16.8 and 23.6 + 7.1, respectively). The abnormal levels of expression of HLA-class II molecules could be related to the altered immune functions observed in elderly people.

HLA antigens associated with sarcoidosis.

Abstract: In a series of 123 sarcoidosis patients (inhabitants of Moravia), frequencies of 15 HLA-A, 31 HLA-B, and 7 HLA-C antigens have been found. (Control group consisted of 500 healthy persons from the same region.) A subgroup of 46 patients was examined in order to determine a frequency of 10 HLA-DR antigens. (Control group consisted of 146 persons.) A positive association was proved between sarcoidosis and the HLA-B8 and B13 antigens (RR = 2.8 and RR = 3.1, respectively). A frequency of B8B13 heterozygotes was highly significant (RR = 8.5). The observed antigen genotype frequencies may be explained by the hypothesis of 2 HLA-linked disposing genes.

Independent mutations in the flanking sequence of the alpha-1-antitrypsin gene are associated with chronic obstructive airways disease.

Abstract: Two restriction fragment length polymorphisms (RFLPs) of the flanking sequence of the alpha 1-antitrypsin (AAT) gene, detected with the restriction enzymes HindIII and TaqI, have been reported to occur more commonly in patients with chronic obstructive airways disease (COAD). Their frequencies were investigated in 20 Caucasian families with a family history of COAD and in 140 unrelated COAD patients none of whom had AAT deficiency. The HindIII polymorphism was present in six index cases of 20 families (p = 0.0015) and 14 of the unrelated patients (p = 0.061) compared with one of 60 healthy unrelated controls. The TaqI polymorphism was present in five of 101 healthy unrelated controls and in three index cases of the 20 families. In the unrelated patient group 28 of 140 had the polymorphism (chi 2 = 10.01, p = 0.0016) and corresponds to a mean log odds ratio of 1.56 (95 per cent confidence limits of 0.58-2.56). The polymorphisms occurred independently of each other and were not associated with AAT deficiency in the basal state.

Molecular diagnosis of the Philadelphia chromosome in chronic myelogenous and acute lymphoblastic leukemias by PCR.

Abstract: We have developed a very rapid method for the detection of the Philadelphia chromosome based on the polymerase chain reaction. This chromosomal translocation generates bcr/abl fusion mRNAs, which were transcribed in cDNAs and subsequently amplified with two sets of primers. Through the use of an additional internal primer set the method reaches the sensitivity to detect a single Ph1-positive cell among 10(5) unaffected cells without any blotting procedures. The whole analytical procedure starting from RNA isolation up to agarose gel electrophoresis can be performed in about 5 h. First results indicate that the frequency of the Ph1 translocation exceeds 40 per cent in certain immunologic subtypes of adult acute leukemia.

Fine needle aspiration cytology : utilization in pediatric pathology

Abstract: Fine needle aspiration can serve as the initial diagnostic modality for a wide variety of lesions within the pediatric age group. The utility of the technique depends on the clinical setting and histologic type of tumor under study. FNA is most valuable for staging and conformation of metastatic spread in small round cell malignancies. The use of aspiration cytology as the initial diagnostic procedure for these neoplasms is more controversial since this technique may deprive the clinician of valuable information (histologic subtype, oncogene status) now available only by examination of large tissue biopsies. Similarly, FNA can serve as a triage technique for the separation of patients harboring thyroid nodules or enlarged lymph nodes into operative candidates and non-operative candidates. As cytopathologists become more familiar with the appearance of pediatric neoplasms, this diagnostic technique will become more widely used, reducing the need for operative intervention in the diagnosis of many benign and reactive lesions.

A novel non-lineage antigen on human leucocytes: characterization with two CD-48 monoclonal antibodies.

Abstract: Two murine monoclonal antibodies have been produced which identify a novel surface antigen expressed on human leucocytes in a non-lineage-restricted distribution. Antibodies WM-63 and WM-68 were derived after immunization of mice with human T-CLL cells and the leukaemic cell line HSB-2. Both antibodies were shown to react with over 90 per cent of normal T and B lymphocytes from peripheral blood and tonsil, and also with monocytes from peripheral blood. A subset of bone marrow leucocytes, including granulocyte-macrophage progenitors, were also reactive. No activity with non-haemopoietic cells or tissues could be identified, however WM-63 and WM-68 showed binding to virtually all cases of chronic B cell malignancy, including chronic lymphatic leukaemia and non-Hodgkin's lymphoma, as well as a proportion of cases of acute leukaemia. Although the antigen recognized by these antibodies could not be immunoprecipitated from membrane extracts, it was removed from the surface of intact cells using the proteolytic enzymes protease and papain. Re-expression on cultured cells was inhibited by incubation with puromycin, cycloheximide, and tunicamycin, indicating that the epitopes detected by WM-63 and WM-68 are likely to be carbohydrate moieties on a protein backbone. Removal of the antigen from the cell surface by treatment with the enzyme phosphatidyl-inositol phospholipase C indicates that it is linked by a phosphatidyl-inositol bond. WM-63 and WM-68 were both recently clustered at the Fourth International Workshop on Human Leucocyte Differentiation Antigens into CD-48, together with four other monoclonal antibodies. Although no biological function has been ascribed to the molecule detected by these antibodies, its restriction to the haemopoietic lineage suggests a role in regulation of leucocyte function.

HLA and celiac disease in Argentina: involvement of the DQ subregion.

Abstract: A population of 62 unrelated homogeneous Argentinian celiac pediatric patients were typed for HLA-A,B,C,DR, and DQ antigens. The association between celiac disease and the DR3 and DR7 antigens was confirmed. The specificity DQw2 was present in 95.2 per cent of the patients. Nevertheless, it was of interest that the most significant phenotypes observed were DR3/DR7, DR7/DR5, and DR3/DR5. The significance of these findings is discussed.

Serological and molecular studies of HLA in insulin-dependent diabetes mellitus in Sardinia.

Abstract: This study was carried out in Sardinia, an Italian region with a very high IDDM incidence. HLA class I and class II antigens were studied in 97 unrelated IDDM patients, 33 complete families with at least one affected member each, and 559 healthy controls. Molecular typing of the DQB1 alleles was carried out in 31 patients and 61 controls. The haplotypes were determined by family studies. The HLA-DR3, DQw2, and DR4 antigens were positively associated with IDDM. The DR3 antigen was nearly always associated to B18 and frequently carried by the extended haplotype A30 Cw5 B18 3F130 DR3 DQw2. The genotype analysis of the patients showed a strong increase of the DR3/DR4 heterozygotes with a relative risk higher than that of the DR3 and DR4 homozygotes. The DR2 antigen was negatively associated with IDDM in the central island districts but not in the southern districts. The DQB1 molecular analysis showed only three alleles in the patients: DQB1*0201 (75.8 per cent), DQB1*0302 (16.1 per cent), and DQB1*0502 (8.1 per cent). These alleles are non Asp 57, so it would seem that nearly if not all Sardinian IDDM patients are NA/NA homozygotes. The DQB1*0502 allele, extremely rare in other Caucasian populations, represents in Sardinia about 70 per cent of the HLA-DR2 haplotypes, contributing to the increase of the pool of IDDM susceptible genes. Moreover it is carried in 27 per cent of the DR2 positive individuals with the extended haplotype A2 Cw7 Bw58 3F31 DR2 DQw1.AZH.

HLA-DP antigens in HIV-infected individuals.

Abstract: We studied the distribution of HLA-DP antigens in 74 HIV-infected Danish homosexual men and 188 ethnically matched healthy individuals, using the primed lymphocyte typing (PLT) technique. Forty of the patients developed AIDS within 3 years after diagnosis, whereas the remaining 34 were healthy or had only minor symptoms for 3 years or more (median observation time was 42 months). HLA-DPwl seemed to be decreased (relative risk = 0.3) in AIDS patients (5.0 per cent) when compared to patients with minor symptoms (14.7 per cent) and healthy controls (14.9 per cent). These differences were, however, not statistically significant. There were no other apparent deviations between patients (or subgroups of patients) and controls.

Evidence of HLA-linked susceptibility gene(s) in primary congenital glaucoma.

Abstract: The study included 82 individuals. Twenty-two were sporadic unrelated patients with Primary Congenital Glaucoma (PCG), and 60 were the members of 10 multiplex families. They were enrolled into two groups; GI:10 multiplex families; GII:32 unrelated patients with PCG (22 sporadic plus 10 probands of the multiplex families). The following were carried out for all the individuals: (1) detailed history; (2) ophthalmologic examination for diagnosis of PCG; (3) clinical examination to exclude any other disease; (4) HLA antigen typing using 30 antigens, 9 for A locus, 15 for B locus, and 6 for DR locus. Genetic and statistical analysis revealed the following: (1) significant association between HLA-B8 and PCG; (2) family studies revealed a susceptibility gene, probably recessive, predisposing to PCG; (3) this gene is linked to HLA, with strong linkage disequilibrium with B8.

Non-HLA genetic factors and insulin dependent diabetes mellitus in the Japanese: TCRA, TCRB and TCRG, INS, THY1, CD3D and ETS1.

Abstract: Polymorphism of the genes encoding the alpha, beta, and gamma chains of the human T-cell receptor (TCRA, TCRB, and TCRG), insulin gene (INS), and three closely linked polymorphic genes on chromosome 11q23, Thy-1 (THY1), T3-D (CD3D), and c-ets proto oncogene (ETS1) were investigated among 56 unrelated patients with insulin-dependent diabetes mellitus (IDDM) and 48 healthy controls. Only eight of the 17 enzymes examined revealed restriction fragment length polymorphism (RFLP), with the use of TCRA, TCRB, and TCRG. No significant association was observed. Polymorphism after BglI, SstI, and TaqI digestion was observed for the INS gene. In consideration of the three classes within the insulin-gene-linked DNA polymorphism alleles, A1 and more rarely A2 alleles were found, but with no significant frequencies. THY1 and CD3D genes were polymorphic after MspI digestion but no significant association was observed. Conversely, the ETS1 gene showed polymorphism after TaqI, SstI, and AvaII were used. Only a significant AvaII-polymorphic fragment (p less than 0.03) was found. However, this significant association disappeared when the correct p value was applied. These results were compared to findings in Caucasians and some differences were noted. The polymorphism observed in this study may be useful in genetic studies on immunologically affected populations.

Histocompatibility antigens (A, B, C, and DR) in Arabs with rheumatoid arthritis.

Abstract: Eighty-five Arab patients in Kuwait with classical and definite rheumatoid arthritis were typed to examine the frequency of HLA-A, B, C, and DR antigens. The results showed a significant increase in the frequency of HLA-A10, B8, B21, and DR3 antigens when compared to an age- and sex-matched control population. HLA-DR3 was present in 34 per cent of the patients compared with 2 per cent of the controls (p less than 0.001). The association of RA in the Arab population with HLA-DR3 rather than HLA-DR4 or HLA-DR1 as reported in other ethnic groups emphasizes further the complexity of the genetics of RA.

HLA-DRw15 is increased in frequency in Japanese scleroderma patients.

Abstract: HLA-DRB allogenotypes were compared in 18 Japanese scleroderma patients and healthy Japanese controls. HLA-DRw15 was found to be significantly increased in frequency in the patient series compared to controls (chi 2 = 4.25, p less than 0.05, Yates' corrected); in DRw15 positive individuals the relative risk of developing scleroderma was 4.0. In Caucasoids DRw11 is significantly associated with scleroderma (Dunckley et al., 1989) and sequence data shows that DRw11 and DRw15 DRB molecules (together with DRw8 where the relative risk of developing scleroderma in Caucasoids is 2.1) share the same amino acid sequence at position 67-70 in the first domain. This would suggest that the DRB1 locus may well be the primary disease promoting locus in scleroderma.

Analysis of CD1 molecules on haematological malignancies of myeloid and lymphoid origin. I. Cell surface antigen expression.

Abstract: One hundred and ninety well-characterized acute and chronic leukaemias were studied for the expression of CD1a antigen by indirect immunofluorescence (IIF). CD1a was detected on 28 per cent of mature B cell lymphoproliferative disorders, 26 per cent of acute non-lymphoblastic leukaemias (ANLL), 21 per cent of chronic granulocytic leukaemias in blast crisis (CML-BC), 53 percent of T acute lymphocytic leukaemias (T-ALL) and in only one out of 35 common acute lymphoblastic leukaemias (c-ALL). In some cases the expression of the CD1a antigen on the surface of leukaemic cells showed a spontaneous fluctuation after a short period of incubation in vitro. CD1b and CD1c molecules were also detected on B cells and acute non-lymphoblastic leukaemias. The presence of CD1 antigens was confirmed using a dot blot assay (DBA) on the lysate of leukaemic cells.

HLA-antigens in a Tunisian familial chondrocalcinosis.

Abstract: Thirty members of a Tunisian family with hereditary chondrocalcinosis were typed for HLA-A, B, and DR antigens: 7 affected and 23 unaffected subjects in three consecutive generations. The haplotype A1 B12 DR3 was found in all affected subjects and in 8 unaffected members. Chondrocalcinosis in this family may be associated with the haplotype A1 B12 DR3. The mode of transmission was autosomal dominant with incomplete penetrance.

Southern blot analysis of DNA extracted from paraffin wax-embedded tissue.

Abstract: Genetic analysis of multi-generation families using RFLP is often incomplete because fresh tissue is not available for the isolation of DNA. However, in many instances archival material exists, usually formalin-fixed in paraffin blocks. Procedures have been described for extracting DNA from such specimens. Many parameters affect the quality of the DNA eventually extracted and this in turn determines whether the DNA can be successfully digested with restriction enzymes and probed (Dubeau et al., 1986; Warford et al., 1988). In essence, the procedures followed at the time of fixation largely determine whether archival material will yield useful results. We have re-examined the effect of varying the fixation time but, more importantly, we have assessed the validity of RFLP results obtained from fixed tissue.

Genetic epidemiology of non-insulin-dependent diabetes mellitus (NIDDM) in north India: distribution of Gm and Km allotypes in 'Punjabis'.

Abstract: Non-insulin dependent diabetes mellitus (NIDDM) is becoming endemic to modernizing and urbanizing populations all over the globe. The data on Gm and Km allotypes in the 'Punjabis' (n = 165) from north India show an interesting association with NIDDM. Differences between diabetic and control samples for the Km allele frequency distribution approach significance (p = 0.0897). The observed phenotypic diversity in the Gm system in diabetics indicates genetic heterogeneity in the disease group. A significant decrease (or absence) of phenotype Gm1,3,17; ;5,21 (p = 0.0119), and presence of phenotype Gm1,2,3,17; ;5,21 (p = 0.0158) in diabetics may constitute a risk factor for this disorder in 'Punjabis'.

DNA variants of alpha-1-antitrypsin in rheumatoid arthritis with and without pulmonary complications.

Abstract: Restriction fragment length polymorphisms (RFLPs) of alpha-1-antitrypsin were studied in 99 subjects with rheumatoid arthritis alone, 21 subjects with rheumatoid arthritis and pulmonary fibrosis, 26 subjects with rheumatoid arthritis and bronchiectasis, and 86 controls. No associations with either rheumatoid arthritis itself or with the associated pulmonary disorders were noted in this U.K. Caucasoid population.

Extended HLA haplotypes in multiplex families with insulin dependent diabetes mellitus in Tunisian population: HLA serological typing and RFLP analysis.

Abstract: Haplotypes including HLA A, B, C, DR, and DQ were compared in a study population comprising 18 Tunisian multiplex families with diabetic children. Eighty haplotypes found in IDDM patients were compared with 148 haplotypes present in healthy family members. RFLP analysis showed that two DR subtypes were significantly more common in the diabetic haplotypes (DR4-DQw8: 82 per cent in IDDM members compared to 0 per cent in healthy members, p less than 0.001 and DR-Dw25: 56 per cent in IDDM patients compared to 16.7 per cent in healthy members, p less than 0.001) and these were in most cases found in haplotype combinations with HLA A2 B44 DR4 DRw53 and HLA A 24 B18 DR3 genes, respectively.

Phenotypic abnormality of T cells in B cell non-Hodgkin's lymphoma.

Abstract: CD4+ T cells of patients with B cell non-Hodgkin's lymphoma (NHL) were analysed for expression of CD45RA and CD29. It was found that CD45RA expression was significantly lower, and CD29 expression significantly higher, in lymphoma patients compared to normal controls. Moreover absolute numbers of CD4+ T cells were significantly lower in NHL patients, due to selective depletion of CD4+ CD45RA+ cells.

HLA antigens in Greek patients with cholelithiasis.

Abstract: The distribution of HLA-A and B antigens was studied in 100 patients with cholelithiasis and in 202 healthy individuals all of Greek origin. An increased frequency of HLA-Aw19 was found in patients (33 per cent) compared with controls (22 per cent) (p less than 0.05, RR 1.7). The increase of Aw19 was even higher for patients with a family history of the disease (44 per cent, vs 22 per cent, p less than 0.01, RR 2.7) and for patients with cholesterol gall stones (44 per cent vs 22 per cent, p less than 0.01, RR 2.5). No difference was seen in patients without a family history of disease or with mixed gall stones. These results suggest a genetic basis for the development of cholesterol gall stone disease.

Elevated c-myc messenger RNA in multiple myeloma cell lines.

Abstract: Oncogene analyses of four human myeloma cell lines provided no indication of gene amplification or rearrangement using DNA probes for the met, raf, abl, mos, erb B, Her-2-neu, fos, myb-7, fms, L-myc, sis, and myb-1 genes. However, a consistent elevation of up to 23-fold in the level of c-myc mRNA was observed in all of the cell lines studied. No restriction fragment length polymorphism (in exons one, two, or three) or c-myc gene amplification has as yet been demonstrated to account for the c-myc mRNA elevation. The c-myc mRNA has a half-life of 25 min which is comparable to that observed in other systems. The elevation in c-myc mRNA is further evidence for the role of the c-myc proto-oncogene in the pathogenesis of myeloma.

B cell differentiation and lymphocyte surface phenotype in late onset hypogammaglobulinaemia.

Abstract: Lymphocyte function and cell surface phenotype were examined in fifteen patients with late onset hypogammaglobulinaemia. The percentage of surface immunoglobulin-positive B cells in fourteen of the fifteen patients was in the normal range. Patients' B cells expressed MHC class II antigens at normal levels. For one patient, there was relatively high sIgD and low sIgM expression on B cells; the rest of the patients did not differ from controls in surface immunoglobulin density. The proportion of B cells positive for CD5 in patients was comparable to normal controls, and considerably less than in cord blood. However, the pattern of immunoglobulin isotype secretion in vitro by patients' B cells closely paralleled responses of cord blood B cells. Spontaneous secretion of IgM and IgG by patients' B cells was very low. Following polyclonal activation in the presence of autologous T cells, cells from thirteen patients secreted IgM within the normal range in response to at least one activator. The response of patients' purified B cells to IL-2 and gamma-IFN was variable. For four of six tested, B cells cultured with IL-2 and gamma-IFN together with polyclonal activators secreted normal levels of IgM. B cells from the other two patients secreted little or no IgM in response to these cytokines. For fourteen patients, IgG secretion following polyclonal activation remained low both when B cells were cultured with T cells or with a combination of IL-2 and gamma-IFN. IgG subclass imbalance was seen in one patient, whose cells secreted an unusually high proportion of IgG3, and undetectable IgG2 and IgG4; this pattern was consistent whether T cell help was provided by autologous or allogeneic T cells. Similarly purified B cells from this patient showed deficient IgG2 and IgG4 production in response to IL-2 and gamma-IFN.

Analysis of CD1 molecules on haematological malignancies of myeloid and lymphoid origin. II. Intracellular detection of CD1 antigens.

Abstract: The surface and cytoplasmic expression of CD1a molecules was analysed by indirect immunofluorescence (IIF) and dot blot assay (DBA) in a panel of 40 acute and chronic leukaemias. Thirty-two per cent of the samples were positive by IIF but, surprisingly, 72 per cent of the patients were positive by DBA, suggesting the intracellular presence of these molecules, CD1b and CD1c were also detected by DBA at similar percentages. Immunocytochemical staining of cytocentrifuge preparations confirmed the intracellular presence of CD1a, CD1b, and CD1c in leukaemic cells of pre-B, B, T, and non-lymphoid lineages.

Immunoregulatory T cells in B cell chronic lymphocytic leukaemia: evidence of a unique phenotype a flow cytometric study.

Abstract: CD4+ T cells of 57 patients with B cell chronic lymphocytic leukaemia (CLL) were analysed for expression of CD45RA and CD29. The majority of CLL patients (33 cases) showed a novel coexpression of these markers on a significant proportion of CD4+ T cells; however, analysis of a single blood sample revealed no apparent prognostic value associated with the markers. Expression of CD45RA and CD29 correlated with parameters of immune function consistent with the known attributes of these markers.

CAR-3 and CA 19-9 serum levels in pancreatic cancer: any differences between two epitopes of the same mucin-like glycoprotein?

Abstract: The aim of this study was to compare the utility of two recently identified tumour markers of pancreatic cancer, CA 19-9 and CAR-3, and to ascertain the roles of some factors influencing both antigens CA 19-9 and CAR-3 were measured in sera of 18 control subjects, 27 patients with pancreatic cancer, 25 with chronic pancreatitis, and 29 with extra-pancreatic diseases CA 19-9 and CAR-3 were, respectively, found to be increased in 85 per cent and 44 per cent of patients with pancreatic cancer, 28 per cent and 0 per cent with chronic pancreatitis and 72 per cent and 28 per cent with extra-pancreatic diseases The ROC curves showed that, for any serum value considered, CA 19-9 is more effective than CAR-3 in discriminating between pancreatic cancer and control subjects and chronic pancreatitis With the combined use of both antigens the results were no better than those given by CA 19-9 alone Correlations were found between liver function tests and CA 19-9 levels and between cholestasis indices only and CAR-3 values Our findings show that CAR-3 is not a sufficiently reliable marker of pancreatic cancer, due to its low sensitivity Nor does it offer any more information than CA 19-9 Both assays are influenced, at least in part, by the extent of the neoplasia Cholestasis which can greatly influence a serum glycoproteic marker such as CA 19-9, was found also to affect, to a lesser extent, CAR-3, an epitope on the same mucin molecule

Xeroderma pigmentosum: clonal chromosomal rearrangements in a pre-cancerous skin lesion.

Abstract: In cells from a papulonodular formation of a patient with the clinical and cellular phenotype of the variant form of xeroderma pigmentosum (XP-V), clonal rearrangements involving different chromosomes were observed. This finding confirms the literature data suggesting that multiple non-specific chromosome anomalies are typical of pre-malignant and malignant skin lesions.