Inhibition by adenine of in vitro immunological functions of normal and adenine phosphoribosyltransferase-deficient human lymphocytes.

Exogenous adenine strongly inhibited mitogen-stimulated transformation, cytoplasmic immunoglobulin production, and natural killer activity of human mononuclear leukocytes at the high concentration of 1.0 mM. These inhibitions by adenine were not due to cytotoxicity, because the viability of cultured cells was not affected by adenine up to 1.0 mM. As the magnitude of inhibition by adenine of these in vitro immunological functions was similar in normal and adenine phosphoribosyltransferase-deficient cells, its inhibition was not mediated by corresponding nucleotides. Adenine at the concentration of 0.1 mM caused 50% inhibition of cytoplasmic immunoglobulin production without alternating cell proliferation or viability. This suggests that an appropriate concentration of adenine may inhibit the differentiation of B cells to plasma cells rather than affecting cell proliferation. Understanding the mechanisms of adenine inhibition may lead to new approaches for the regulation of immune responses.

Natural killer activity in two cases of ataxia telangiectasia.

We report two cases of ataxia telangiectasia in which a small number of T cells and a slightly diminished response to phytohaemagglutinin or concanavalin A were observed. In one case, serum IgA was undetectable. In spite of T and/or B cell involvement, natural killer activity and the percentage of natural killer cells determined by monoclonal antibodies Leu 7 were normal in both cases.

LPF-induced T cell colony formation: effect of PMA and interleukin-2, and surface marker analysis.

We demonstrated that a primary exposure to the lymphocytosis promoting factor (LPF) of Bordetella pertussis-induced T cell colony formation. Colony formation was observed when mononuclear cells (MNC) were cultured at concentrations of more than 1 X 10(6)/ml, and reached a peak on day 8. However, the number of colonies generated with LPF was about one-third induced with phytohaemagglutinin (PHA). Removal of monocytes from MNC or T cells resulted in the failure of colony formation, but colony growth could be restored by the addition of monocytes or B enriched cells, indicating that they were required for the optimal colony growth induced by LPF. In the absence of accessory cells, optimal colony growth from monocyte depleted T cells could be obtained when an appropriate concentration of phorbol myristate acetate (PMA) or interleukin-2 (IL-2) was added in the cultures with LPF. PMA did not enhance LPF-induced colony formation in the cultures containing a sufficient amount of exogeneous IL-2. These findings suggest that IL-2 is essential to LPF-induced colony formation. Surface marker analysis showed that most of LPF-induced colony cells were T cells. The percentages of T4+ and T gamma cells of LPF-induced colony cells were more, and T8+ cells less, than those of PHA-induced colony cells. Ia1, T9 and Tac antigens were detected on many colony cells induced by LPF or PHA. These results indicate that the phenotype of LPF-induced colony cells differs from those of PHA, but the sequential antigen expression on lymphocytes triggered by IL-2 might be similar in both LPF- and PHA-induced colony formation.

The mitogenic effect of the lymphocytosis promoting factor from Bordetella pertussis on human T gamma and non-T gamma cells.

We studied the effect of lymphocytosis promoting factor (LPF), derived from the supernatant fluid of a culture of phase I Bordetella pertussis strain Tohama, on human lymphocyte proliferation. LPF was a potent mitogen for human mononuclear cells, specifically T cells. LPF failed to induce cytoplasmic immunoglobulin production by B cells. Removal of the monocytes from the T cell fraction diminished responses to LPF, but the response could be restored completely by the addition of 5.0% monocytes. These results suggest that LPF-induced cell proliferation is at least partially dependent on monocytes. In contrast to PHA, LPF stimulated T gamma cells to a greater extent than non-T gamma cells, but the magnitude of the T gamma or non-T gamma cell response was less than that of T cells, indicating that synergistic interactions between T gamma and non-T gamma cells are required for maximal response.

Comparison of phorbol myristate acetate and phytohaemagglutinin as stimulators of in vitro T lymphocyte colony formation of human peripheral blood lymphocytes. I. Surface markers of colony cells.

Human T lymphocyte colonies were grown in methylcellulose semi-solid cultures in the presence of phytohaemagglutinin (PHA) and/or phorbol myristate acetate (PMA). Surface marker analysis showed lower percentages of OKT3- and OKT4-positive cells in PMA-induced colonies than those in PHA-induced colonies. The percentage of OKIa1-positive cells in PMA-induced colonies was approximately twice that in PHA-induced colonies. The percentage of OKT9-positive cells in PMA-induced colonies was significantly lower than that in PHA-induced colonies. These data suggest that the subsets of PMA-induced colony cells express a more immature phenotype than that of PHA-induced colony cells and that, among PMA-induced colony cells, there are fewer T cells in the proliferative status at the time tested. When 3 X 10(5)/ml monocyte-depleted T cells, at which concentration of seeded cells neither PHA nor PMA could induce colony growth, were cultured in the presence of both PHA and PMA, T cell colony growth was observed. In T cell colonies induced by a combination of PHA and PMA, the percentages of OKT3-, OKT4- and OKT8-positive cells were different from those in colonies induced by either PHA or PMA alone. These results suggest that PMA acts not only as a substitute for monocytes and/or interleukin-1, but may directly affect lymphocyte proliferation induced by a combination of PHA and PMA.

Effects of tunicamycin and various monosaccharides on phytohaemagglutinin-induced autorosette formation.

The structural characteristics of autologous red blood cell (ARBC) receptors on human peripheral blood lymphocytes (PBL) induced by phytohaemagglutinin (PHA) were examined. Tunicamycin, which is known to be a blocker of the protein glycosylation of N-glycosidic type glycoprotein, significantly inhibited the production of ARBC receptors. When trypsinized PBL were cultured in the presence of tunicamycin, the inhibitory effect was enhanced. When several monosaccharides were added to the mixture of ARBC and PBL pre-treated with PHA for 24 h, D-mannose caused the most inhibition, and galactose and D-fucose also caused significant inhibition. These data suggest that N-glycosidic type glycoproteins play an important role in binding sites to ARBC on PBL surface membranes and that more than one type of glycose may be involved.

Favorable effect of dimethyl sulfoxide on secondary amyloidosis in juvenile rheumatoid arthritis.

A girl with secondary amyloidosis as a complication of juvenile rheumatoid arthritis was administered dimethyl sulfoxide by topical application to the skin. Her gastrointestinal symptoms and massive proteinuria improved. Decreased left ventricular function and creatinine clearance also improved remarkably. The favorable effect of dimethyl sulfoxide in this single patient deserves further study in a controlled trial.

Complete deficiency of adenine phosphoribosyltransferase: a report of three cases and immunologic and phagocytic investigations.

The levels of adenine phosphoribosyltransferase (APRT:EC 2.4.2.7) were determined in red blood cells (RBCs), peripheral mononuclear cells (MNCs), and polymorphonuclear leukocytes (PMNLs) from normal controls and from two families with APRT deficiency. No APRT activity was demonstrated in MNCs and PMNLs of patients with complete deficiency of RBC-APRT. APRT deficiency occurs not only in RBCs but also in MNCs and PMNLs. Immunologic and phagocytic examinations showed normal hemogram and serum immunoglobulin levels, and normal E-rosette forming cells and surface immunoglobulin-bearing cells. Lymphocyte blastogenesis in response to phytohemagglutinin and lymphocyte differentiation to cytoplasmic immunoglobulin-producing cells induced by pokeweed mitogen were normal. No major defects were apparent in natural killer activity. Phagocytic functions were normal as tested by bactericidal activity, O2-consumption, chemotaxis, and chemiluminescence response.

Induction of human T-lymphocyte colonies by phorbol myristate acetate.

Colony growth of human lymphocytes by phorbol myristate acetate (PMA) was studied. PMA was able to induce lymphocyte colony growth in methylcellulose semi-solid cultures in the absence of lectin mitogens, phytohaemagglutinin or concanavalin A. PMA-induced colonies were found in cultures of mononuclear cells, monocyte-depleted mononuclear cells and the T-enriched cell fraction, whereas no colonies were obtained from the non-T cell fraction. At least one million mononuclear cells were required to form colonies by PMA. The colony cells were mainly T cells as judged from sheep red blood cell rosette formation. Surface immunoglobulin positive cells and peroxidase positive cells were not detected in colony cells. Non-specific esterase positive cells were only found to be less than 1% of colony cells. T cells formed more colonies than did mononuclear cells, presumably because of a concentration of colony forming cells and/or co-operating cells. Colony formation by PMA was induced from monocyte-depleted mononuclear cells and monocyte-depleted T cells, suggesting independence of monocytes. When mononuclear cells were precultured at 37 degrees for 5 min or 30 min with PMA and then cultured in the semi-solid medium without PMA, only a small number of colonies grew. Further studies of PMA-induced T-cell colonies will provide information the identification and characterization of immunological states in various immunological diseases.

A case of immunodeficiency with decreased serum IgA levels and impaired polyclonal B cell differentiation in vitro.

We report a patient with low serum IgA levels and persistent pulmonary infection. In spite of the normal contents of serum IgG and IgM, the patient had a deficiency for plasma cell differentiation of all major classes of immunoglobulins (IgG, IgA and IgM). Cross culture systems between normal T or B cells and the patient's T or B cells showed a defect of both T and B cell function.

Inhibition of phytohaemagglutinin-induced autorosette formation of human peripheral blood lymphocytes by RNA or protein synthesis inhibitor.

The induction of autorosette forming cells (ARFC) with phytohaemagglutinin (PHA) in human peripheral blood lymphocytes (PBL) was examined. After 24 h of incubation with PHA, the level of ARFC in PBL was markedly increased but subsequently decreased to about one-third of the peak level at 96 h of culture. When PBL were pre-treated with actinomycin D, or cultured with puromycin, the induction of ARFC was completely blocked. Pre-treatment of PBL with mitomycin C (MMC) had no effect on induction of ARFC, whereas DNA synthesis was completely blocked. These data indicate that the generation of autologous red blood cell receptors is a relatively early event in PHA activated PBL, and that it is independent of DNA synthesis but dependent on RNA and protein synthesis.

Induction of human autorosette forming cells by phorbol myristate acetate.

Human peripheral blood lymphocytes were examined for rosette formation with autologous erythrocytes. When normal human lymphocytes were stimulated with phorbol myristate acetate (PMA) in the presence of autologous serum, the levels of autorosette forming cells (ARFC) were strongly enhanced. Pre-culture was necessary for the generation of ARFC by PMA and the maximal level of ARFC was observed at 72 hr of culture. ARFC appear to belong to a T cell subset and the induction of ARFC by PMA was noted in monocyte depleted lymphocyte fractions, indicating monocyte independency.

Vaccine-associated poliomyelitis in an infant with agammaglobulinemia.

We describe a female infant with agammaglobulinemia who contracted vaccine-associated poliomyelitis. Poliovirus type 2 was isolated from the initial stool specimen. In our patient, temporary changes in the cerebrospinal fluid resembled those in patients without immunodeficiencies, although gammaglobulin therapy had not yet been started. Pleocytosis was observed for a short time after viremia, but soon there was a return to normal without antibody production.