The HLA-DR beta 16 allogenotype constitutes a risk factor for hypertrophic scarring.

Nineteen patients that had developed hypertrophic scars subsequent to thermal injury were typed for HLA class II allogenotypes with the restriction fragment length polymorphism technique. A significant association was found with DR beta 16 (pc = 1.45 x 10(-4); relative risk = 12.25). This finding adds evidence to other data suggesting that immunologic phenomena are involved in pathologic scarring. Moreover, the results presented here have allowed an identification of a genetically determined risk factor for hypertrophic scar formation located in the HLA region.

Gamma-IFN induces a differential expression of HLA-DR, DQ and DP antigens on peripheral blood myeloid leukemic blasts at various stages of differentiation.

Using specific monoclonal antibodies and the indirect immunofluorescence technique, peripheral blood myeloid leukemic blasts from 49 patients were studied for DR expression, 42 for DQ and nine for DP after four days of culture without and with gamma-IFN. The expression of class II molecules on untreated cells depended upon the stage of differentiation and was maximal for DR antigens and lower for DP, while DQ was present only in a small percentage of M2, M4 or M5 blasts. M3 blasts lacked both DR and DQ. Maximal increase of surface expression induced by gamma-IFN was observed for DR molecules independently from the differentiation stage. No significant modification was seen for DQ. Preliminary data concerning DP indicate an increased expression in some of the tested samples. Thus the results obtained on peripheral blood blasts parallel previous observations on leukemic cell lines.

Altered biosynthesis of tumour necrosis factor (TNF) alpha is involved in postburn hypertrophic scars.

The present study shows that the decrease of TNF alpha in postburn hypertrophic scars is due to a decrease in the steady-state level of TNF alpha mRNA and thus to an altered biosynthesis of the cytokine. Thirteen scars, including seven hypertrophic and six normotrophic scars, were tested for TNF alpha mRNA production by a semiquantitative reverse polymerase chain reaction (PCR) method. TNF beta and beta actin were tested as a control. Six out of six normotrophic scar samples amplified with primers for TNF alpha showed a positive PCR signal up to the 1:32 dilution. On the contrary all the hypertrophic tested samples (7/7) had a positive PCR signal only at the 1:1 or 1:2 dilution. All samples, both normotrophic and hypertrophic, were homogeneous as to TNF beta production.

LDLR, GYPA, HBGG, D7S8 and GC allele and genotype frequencies in the northwest Italian population.

Allele and genotype frequencies for five PCR-based DNA markers (LDLR, GYPA, HBGG, D7S8 and GC) were determined in 100 unrelated individuals from Piedmont (Northwest Italy). All five Ioci met Hardy-Weinberg expectations in the sampled population. The combined PD and CE were, respectively, 0.995 and 0.697. Frequencies obtained were compared with other previously published data on Caucasian populations with no significant differences. The genetic data from this study, in addition to those already collected by other groups, contribute to the expansion of the Italian DNA database suitable for forensic casework and paternity testing.

Loss of surface HLA class I molecules in leukemic myeloblasts is correlated with an increased leukocyte concentration at onset.

BACKGROUND: Reduced expression of HLA class I molecules has been demonstrated for many human neoplasias and is correlated with increased malignancy. METHODS: We investigated the correlation between HLA expression on blasts and the number of peripheral blood leukocytes (WBC) at onset in 34 patients with acute non lymphoblastic (ANLL) leukemia. Leukemic blasts at onset were serologically typed for HLA-A,B specificities, and the patients were classified in three groups according to the number of detectable surface HLA antigens. RESULTS: The amount of WBC at onset in patients lacking HLA antigens was significantly higher than in patients with no antigenic loss. CONCLUSIONS: A decrease in HLA class I molecules on the cell surface is correlated with a greater expansion of the leukemic clone.

Expression of class I-like alloantigens on leukemic cells is not correlated with the amount of HLA-A,B,C molecules.

The expression of class I-like allospecificities on leukemic blasts is not correlated to the amount of HLA-A,B,C molecules, as measured with MoAb W6/32 and IIF, nor with the quantitative expression of HLA-A molecules evaluated by absorption studies.

Distribution of tumor necrosis factor alleles (NcoI RFLP) and their relationship to HLA haplotypes in an Italian population.

The NcoI RFLP of the tumor necrosis factor (TNF) beta gene was analyzed in a panel of 105 unrelated healthy Italian blood donors. The gene frequencies of the 10.5 kb and 5.5 kb allele were 0.73 and 0.27, respectively. The 5.5 kb band was significantly positively associated with HLA-A1, B8, DR17.1, and C4AQ0, and negatively associated with DR7.2, DQw9 and C4A6, all being specificities which belong to two well-known Caucasoid ancestral haplotypes. When the population was subdivided on the basis of TNF phenotypes, different linkage disequilibria between HLA alleles were detected in the three phenotypic classes. From this analysis it was possible to relate preferential HLA associations, most of which are characteristic of ancestral haplotypes, to TNF polymorphism.

New HLA class I-like alloantigens expressed on blast cells.

New beta 2-microglobulin (beta 2-m)-associated, HLA-linked alloantigens, selectively expressed on phytohaemagglutinin (PHA)-activated lymphocytes, were identified using human alloantisera. Reactivity of the antisera against activated cells correlated with HLA-A2, A10 and A28 specificities. The new alloantigens were undetectable on peripheral blood mononuclear cells, B lymphocytes and platelets using either the lymphocytotoxicity or the absorption techniques, and appeared on lymphocytes upon PHA activation, with time-dependent kinetics. They were found on some, but not all, acute leukaemias of B, T and myeloid origin, being absent from chronic B lymphocytic leukaemias. The presence of the new beta 2-microglobulin-associated allospecificities was not correlated with the quantity of classical class I antigens present, as analysed on different cell types by flow cytometry. Thus, these antigenic determinants appear to be different from the classical HLA class I antigens and could be the human counterpart of the murine Qa system or of the second H-2K gene.

HLA supratypes in an Italian population.

A supratype analysis of a North Italian population was performed, using 16 polymorphisms in the HLA region spanning the HLA-A-DP segment. Fourteen supratypes were identified, mostly corresponding to those found in other Caucasoid populations. The degree of their conservation both within the B-DR/DQ region and in the regions telomeric and centromeric from HLA-A and DP was evaluated and linkage disequilibria among several DR and DP alleles were identified. Notably, the degree of association with DP increased when the DR marker was part of a conserved B-DR/DQ supratype. These data are relevant to the definition of the genetic structure of the population and to the prediction of probabilities of histocompatibility matching between unrelated individuals.

Quantitative expression of HLA class I molecules in acute non-lymphoblastic leukaemia cells.

The present study concerns a panel of 33 acute non lymphoblastic leukaemia (ANLL) patients, previously typed for HLA-A,B serological specificities and including samples with a normal HLA-A,B phenotype (3,4 detected specificities) as well as samples with missing and extra specificities. Samples were analysed at the protein and/or RNA level in order to verify whether the observed typing anomalies were due to a modified quantitative expression of class I molecules. The number of HLA-A,B assigned specificities correlated significantly with the cell surface class I expression detected by indirect immunofluorescence using the monomorphic anti-class I MoAb W6/32 (Spearman rank correlation test, P < 0.01) and with the amount of class I Heavy Chain (HC, P < 0.05) and beta-2-microglobulin (beta 2m, P < 0.05) evaluated by Western blot in whole cell extracts. The RNA analysis suggested a HC-beta 2m coordinated down regulation at the mRNA level in a patient with no assigned HLA-A,B specificities. Another patient with no detectable HLA-A,B specificities showed a low expression selectively of the beta 2m protein. The results reported here demonstrate a heterogenous quantitative HLA class I expression in ANLL blasts, analogous to results reported for solid tumours.

Altered expression of HLA-A,B specificities on acute lymphoid and myeloid leukaemia blasts.

HLA-A,B specificities were analysed on the neoplastic blasts of a panel of 69 lymphoblastic (ALL) and 50 non lymphoblastic (ANLL) acute leukaemias at onset using the standard lymphocytotoxicity technique. Analysis of the number of detected specificities per locus and, when possible, comparison of the results with those obtained on lymphocytes of the same patients during remission revealed many alterations in the expression of A,B specificities including extra specificities both at the HLA-A and -B loci mainly on lymphoblasts and missed specificities mainly at the HLA-B locus on myeloblasts. Lack of A,B antigens was complete in 6.2% of all tested samples (9% of ANLL) and selective for all the products of one locus in 16.8% of all tested samples (27.7% of ANLL). A decrease of class I molecules on the cell surface was evidenced with MoAb W6/32 on blasts missing detectable serological specificities.

HLA class I- like antigen expression on human leukemic cells.

The expression of HLA class I- like molecules was analyzed on human acute and chronic leukemic cells. The presence on leukemic cells of class I- like molecules, absent on the patient's normal lymphocytes, was examined by complement- dependent lymphocytotoxicity using platelet absorbed alloantisera that recognize HLA-linked, 45-12 kd, beta-2-microglobulin associated molecules, selectively expressed on PHA-activated cells. A positive reactivity of the anti- class I- like alloantisera was found in 50% of the acute leukemias (cALL, T-ALL and AML), independently of the lineage of differentiation, while chronic lymphocytic leukemias (B-CLL) were constantly negative. It is suggested that beta-2-microglobulin associated HLA molecules may represent markers of leukemic blast activation and/or maturation state.