Genetic and clinical description of hemochromatosis probands and heterozygotes: evidence that multiple genes linked to the major histocompatibility complex are responsible for hemochromatosis.

We evaluated Alabama hemochromatosis probands (n = 74) and normal control subjects (n = 142) for expression of the hemochromatosis-associated mutations nt 845G-->A (845A; Cys282Tyr) and nt 187C-->G (His63Asp) in a gene linked to the major histocompatibility complex (MHC). We also tabulated parameters of iron metabolism and iron overload in probands and in obligate heterozygote family members of homozygous Cys282Tyr probands. Among probands, 59.4% were Cys282Tyr homozygotes and 20.3% were heterozygotes; 20.3% did not express this mutation. In normal control subjects, 14.7% were heterozygous for the Cys282Tyr mutation; one normal control subject was homozygous for the Cys282Tyr mutation. None (0 of 44) of our Cys282Tyr-homozygous hemochromatosis probands had the His63Asp mutation. Of the Cys282Tyr-heterozygous and -negative probands, the His63Asp mutation occurred in 26.7% (4/15) and 53.3% (8/15), respectively. In normal control subjects, 23.2% were heterozygous for the His63Asp mutation; 2.8% were homozygous. Induction phlebotomy requirements and other manifestations of iron overload were significantly greater in Cys282Tyr homozygotes than among other probands. Cys282Tyr-heterozygous probands had significantly higher values of serum iron parameters than did obligate Cys282Tyr heterozygotes whose values were, on the average, normal. Co-expression of HLA-A3, HLA-B7, and D6S105(8) was significantly more frequent in all subgroups of probands stratified by Cys282Tyr expression than in normal control subjects. These results demonstrate that the severity of iron overload in hemochromatosis is affected significantly by genetic factors. Further, our findings support the hypothesis that one or more MHC-linked genes other than that corresponding to the Cys282Tyr and His63Asp mutations contributes to increased iron absorption and iron overload in hemochromatosis probands.

Difference in effect of single immunosuppressive agents (cyclophosphamide, CCNU, 5-FU) on peripheral blood immune cell parameters and central nervous system immunoglobulin synthesis rate in patients with multiple sclerosis.

Cyclophosphamide (CY), 1-(2-chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) and 5-fluorouracil (5-FU) were given in single course schedules to chronic progressive multiple sclerosis (MS) patients clinically stable for 6 months. The following peripheral immune cellular parameters were measured before, during and after each drug administration: white blood count (WBC), polymorphonuclear count (PMN), lymphocyte count, percentage of T cells, T cell response to phytohaemagglutinin (PHA), percentage of B cells, percentage of cells bearing receptors for the Fc portion of immunoglobulin (% FcR cells), killer (K) cell activity defined by antibody-dependent cellular cytotoxicity (ADCC), and natural killer (NK) cell activity. Central nervous system (CNS) immunoglobulin G (IgG) synthesis was also measured. The patients were followed carefully by both quantitative and qualitative methods for any change in their neurologic condition. Selective reduction in NK activity was observed with CY and 5-FU while no significant alteration was seen in %FcR cells and K activity. CY differed from 5-FU in reducing lymphocyte count and B cell percentage while 5-FU decreased the percentage of T cells. CCNU, but not the other drugs, reduced T cell proliferative response to PHA. In addition, CCNU, which is known to penetrate well into the nervous system, caused a modest reduction in CNS IgG synthesis, while 5-FU had an uncertain effect. Clinically the patients were unchanged or continued to progress in their disability. The results suggest an independence of the CNS immune from the systemic immune system in MS in response to many immunosuppressive drugs.

In vitro depression of human lymphocyte mitogen response (phytohaemagglutinin) by asbestos fibres.

Asbestosis is a fibrotic lung disease associated with chronic inhalation of asbestos dust. The response of peripheral blood mononuclear cells (PBM) to phytohaemagglutinin (PHA) in asbestosis patients has been reported to be impaired, suggesting a disturbance in the cell-mediated response of chronically exposed individuals. We demonstrated that PHA responses of normal PBM are also depressed when exposed to various forms of asbestos fibres in vitro. Furthermore, we showed the primary effect of the fibres to be on lymphoid (non-adherent) populations rather than monocytes (adherent cells). Exposure as brief as 1 hr affected the subsequent PHA response of the cells. This effect did not appear to involve suppressor cell activation nor was it mediated by soluble factors. Our findings therefore offer an explanation for the alterations in the cellular immune response observed in humans as a result of lymphoid cells coming into transient contact with inhaled asbestos fibres residing in the lung.

Analysis of histocompatibility requirements for proliferative and helper T cell activity. T cell populations depleted of alloreactive cells by negative selection.

T cell populations were prepared from donors immunized with hapten-carrier conjugates and were depleted of alloreactive cells by negative selection. This was accomplished by injection of the cells into H-2-disparate irradiated recipients and recovery from the thoracic duct after 18-40 h. The genetic requirements for the proliferative and helper activity of these populations was determined. The proliferative response to antigen presented on adherent, Thy-1-negative cells was determined, and a requirement for syngeneic antigen-presenting cells (APC) was demonstrated. The same T cells were assayed for their ability to give help to hapten primed B cells. It was shown that there was a requirement for syngeneic APC and for linked recognition of hapten and carrier determinants on the same molecule by the B cell and T cell. There was no requirement for the B cell to be H-2 compatible with the T cell. The requirement for linked recognition was taken as evidence that the responses in allogeneic combinations were not a result of positive allogeneic effects. Precisely comparable restrictions were found with positively selected cells.