Cytochrome P450 1A2 is a hepatic autoantigen in autoimmune polyglandular syndrome type 1.

Autoantibodies directed against proteins of the adrenal cortex and the liver were studied in 88 subjects of Sardinian descent, namely six patients with autoimmune polyendocrine syndrome type 1 (APS1), 22 relatives of APS1 patients, 40 controls with other autoimmune diseases, and 20 healthy controls. Indirect immunofluorescence, using tissue sections of the adrenal cortex, revealed a cytoplasmatic staining pattern in 4 of 6 patients with APS1. Western blotting with adrenal mitochondria identified autoantigens of 54 kDa and 57 kDa, Western blotting with placental mitochondria revealed a 54-kDa autoantigen. The 54-kDa protein was recognized by 4 of 6 patients with APS1 both in placental and adrenal tissue, whereas the 57-kDa protein was detected only by one serum. Using recombinant preparations of cytochrome P450 proteins, the autoantigens were identified as P450 scc and P450 c17. One of six APS1 patients suffered from chronic hepatitis. In this patient, immunofluorescence revealed a centrolobular liver and a proximal renal tubule staining pattern. Western blots using microsomal preparations of human liver revealed a protein band of 52 kDa. The autoantigen was identified as cytochrome P450 1A2 by use of recombinant protein preparations. P450 1A2 represents the first hepatic autoantigen reported in APS1. P450 1A2 usually is not detected by sera of patients with isolated autoimmune liver disease and might be a hepatic marker autoantigen for patients with APS1.

Membrane-bound and soluble IL-2 receptors (p55 and p75 chains) on peripheral blood mononuclear cells from patients with solid malignancies.

The aim of the investigation was to study directly the IL-2 receptor (IL-2R) and its subunits, p55 and p75 chains, either membrane-bound or soluble, on PBMC of patients with solid malignancies and, indirectly, the same patients' PBMC ability to produce IL-2. Fifty-eight cancer patients, 29 men and 29 women, were studied: their mean age was 57.3 yr, range 35-79. Twenty-two healthy age-sex-matched subjects served as controls. The tumors were the most common and the most representative among human cancers, i.e., breast, lung, head and neck, digestive tract and liver, prostate and gynecologic cancers: they were generally in advanced stages and in 23 cases metastatic. The PBMC proliferative response to PHA, PHA plus IL-2, and IL-2 was evaluated along with the response to PHA in the presence of anti-p55, anti-p75 monoclonal antibodies, or both. Moreover, membrane-bound IL-2R (p55 and p75 chains) on PHA-stimulated PBMC was detected, along with soluble IL-2R in the serum and in the culture supernatants. The conclusions suggest that in solid malignancies: the membrane-bound IL-2Rs, both p55 and p75 chains, are expressed normally, there is an high serum level of soluble IL-2R, there is a normal release of soluble IL-2R in culture, and there is an indirect evidence of a lack of IL-2 production. Therefore, no primary impairment of IL-2R was found in solid tumors. Moreover, in our study we have found no difference in any parameter studied between patients with and patients without metastases.

Hodgkin's disease presenting in 1 of 4 siblings affected by hereditary spinocerebellar ataxia: clinical, immunological and genetic study.

One of 4 siblings affected by hereditary spinocerebellar ataxia (HSCA) of Marie's type developed Hodgkin's disease (HD): the stage was IV B, the patient was submitted to conventional chemo- and radiotherapy and achieved complete remission. An accurate clinical, genetic and immunological study was carried out on all his family, including a complete HLA typing, a chromosome study, the immunophenotyping of peripheral blood mononuclear cells (PBMC), the PBMC response to polyclonal mitogens, to interleukin 2 (IL-2), to the association of PHA + IL-2 and the evaluation of the IL-2 receptor expression. No association was clearly demonstrable between an HLA haplotype and HSCA, while the patient with HSCA and HD was HLA-B18- and DQw3-positive (the last at homozygous level), two antigens known to be strongly associated with HD, mainly among the Sardinian ethnic group. The mode of inheritance of HD susceptibility is however completely different from that of Marie's HSCA. The chromosome study did not show any characteristic pattern of the karyotype, neither of the HSCA affected nor of the unaffected members. The immunological investigations did not elucidate any characteristic behavior of the family members, apart from the typical findings of HD seen on patients with HSCA and HD. Our study could not demonstrate any genetic and/or immunologic common background shared by the two diseases, HSCA and HD. Their coexistence in our patient, although the statistic probability is very low, seems to be a fortuitous coincidence more than the result of a common genetic and pathogenetic mechanism.

Immunologic assessment determined by response to IL-2 and immunophenotyping of tumor-infiltrating lymphocytes, of invaded and non invaded lymph nodes and of peripheral blood lymphocytes from twenty-one patients with primary laryngeal cancer.

The aim of the study was to identify the lymphocyte sets and/or subsets possibly involved in the response to malignant cells. For this purpose we have investigated both the cells at the tumor site, i.e. tumor-infiltrating lymphocytes (TIL), and the cells present in the draining lymph nodes, either invaded or non invaded, as well as the peripheral blood lymphocytes from twenty-one patients with primary laryngeal epidermoid carcinoma. The functional assay was carried out by the proliferative response to mitogens, to Interleukin 2 and to their association, the surface immunophenotyping was performed with a large panel of monoclonal antibodies. TIL are the most responsive cells to mitogens, while the responsiveness of TIL and of LN cells to IL-2 was in the same range. PHA-activated TIL are the most responsive cells to IL-2. Our data indicate that TIL do show in vitro, and probably also in vivo, "activation" with elevated responsiveness to IL-2. The surface phenotype showed a strikingly increased proportion of T8+ cells in TIL as compared to T8+ cells in all types of LN, thus confirming within TIL variable, but high, proportions of clones which display cytolytic activity, possibly induced by IL-2. Our data seem to support the perspective for a therapeutic approach in vivo with IL-2, which via its influence on TIL, may act on tumor cells.

Familial chronic B-cell malignancy. Hairy cell leukaemia in mother and daughter.

Two familial cases of hairy cell leukaemia are reported: a daughter, 44-years-old, with a very unusual ultrastructural pattern found in hairy cells, the "tubuloreticular inclusions", and her mother, 71-years-old, who was affected six years later. Routine laboratory investigations, cytochemical and cytogenetic studies including HLA typing, as well as in vitro proliferative response of peripheral blood mononuclear cells (PBMC) to polyclonal mitogens and to exogenous interleukin 2, were performed. The immunological characterization by assessing the cell surface phenotypic markers with monoclonal antibodies and transmission electron microscopy (TEM) investigations were also carried out. In case 2 all tests were performed both on PBMC and on the bone marrow cells. To the best of our knowledge this is one of the first reports of such familial association. The possibility that genetic factors might play a role in the etiology of leukaemia in man is discussed: in our two cases, however, cytogenetic studies did not support this, while HLA typing revealed a non-significant association of HCL with DQw3 allele. Alternatively, an environmental factor has been considered, and a viral infection-perhaps by a retrovirus of the HTLV family has been suggested as tubuloreticular inclusions have been found in both hairy cell leukaemia, as reported, by us, and AIDS-LAS. However, a long time elapsed between the manifestation of HCL in the daughter and in the mother, and as the two patients had not been living together at that time, the possibility of a viral transmission seems minimal. The results of TEM and of immunological investigations are presented and discussed. Both, but particularly the latter, support the B cell nature of the hairy cell.

Hairy cell leukemia with ultrastructural finding of 'tubuloreticular inclusions' in hairy cells: a possible marker of a virus-induced disease?

This report describes a case of hairy cell leukemia (HCL) with typical hematological and clinical findings. The most striking feature is the electron microscopic pattern of intracytoplasmic inclusions within hairy cells (HCs), which can be identified with the 'tubuloreticular inclusions', very unusual and not yet reported in HCL. The same structures were frequently detected in peripheral mononuclear cells from patients with acquired immune deficiency syndrome (AIDS) and with chronic lymphadenopathy syndrome (LAS), that are caused by the HTLV-III retrovirus. In the same patients elevated serum interferon levels were also found. The close relationship between tubuloreticular inclusions, viral infections and serum interferon levels suggests an etiologic association between a virus infection and outbreak of HCL. The results of our laboratory investigations also support a role for interferon in this disease.