Search

Muscle specific tyrosine kinase myasthenia gravis (MuSK MG) is a form of autoimmune MG that predominantly affects women and has unique clinical features, including prominent bulbar weakness, muscle atrophy, and excellent response to therapeutic plasma exchange. Patients with MuSK MG have predominantly IgG4 autoantibodies directed against MuSK on the postsynaptic muscle membrane. Lymphocyte functionality has not been reported in this condition. The goal of this study was to characterize T cell responses in patients with MuSK MG. Intracellular production of IFN-gamma, TNF-alpha, IL-2, IL-17, and IL-21 by CD4+ and CD8+ T cells was measured by polychromatic flow cytometry in peripheral blood samples from 11 Musk MG patients and 10 healthy controls. Only one MuSK MG patient was not receiving immunosuppressive therapy. Regulatory T cells (Treg) were also included in our analysis to determine if changes in T cell function were due to altered Treg frequencies. CD8+ T cells from MuSK MG patients had higher frequencies of polyfunctional responses than controls, and CD4+ T cells had higher IL-2, TNF-alpha, and IL-17. MuSK MG patients had a higher percentage of CD4+ T cells producing combinations of IFN-gamma/IL-2/TNF-gamma, TNF-alpha/IL-2, and IFN-gamma/TNF-alpha. Interestingly, Treg numbers and CD39 expression were not different from control values. MuSK MG patients had increased frequencies of Th1 and Th17 cytokines and were primed for polyfunctional proinflammatory responses that cannot be explained by a defect in CD39 expression or Treg number., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Lymphocytes from HIV-1-seropositive and -seronegative individuals were examined to determine whether HIV-1 infection interfered with the ability to generate a lymphokine-activated killer (LAK) cell response. Following a 3-day ex vivo incubation in the presence of 1000 U/ml of recombinant interleukin-2, lymphocytes from seropositive individuals exhibited a LAK cell response which was equivalent to or greater than that of seronegative controls as measured against Daudi cell targets. LAK cells from seropositive and seronegative donors showed no specific cytolytic activity against gp120-coated or HIV-1-infected targets. However, in the presence of patient sera, significant levels of virus-specific LAK cell-mediated antibody-dependent cellular cytotoxicity (ADCC) were observed. The level of this specific LAK cell-mediated ADCC was greater than that mediated under similar conditions by freshly isolated peripheral blood mononuclear cells. The greatest improvement in ADCC over baseline activity was seen with lymphocytes from AIDS patients after the 3-day ex vivo activation, suggesting that this patient population might benefit the most from adaptive LAK cell therapy., (Copyright 1998 Academic Press.)

Administration of high-titered goat anti-FLV gp71 IgG to AKR mice during a narrow neonatal therapy "window" suppresses the early development of MuLV infectious cell centers (ICC) in spleen, thymus, and bone marrow. By 4-5 weeks of age ICC appear in spleen and marrow cell populations, rapidly increasing to plateau levels within 2 weeks in the absence of viremia. The thymus of treated animals remains devoid of detectable ICC throughout the 4-month period of testing. This pattern of ICC suppression occurs independently of virus neutralization as shown by the inability of F(ab')2 preparations, which retained neutralizing activity, to prevent early establishment of ICC. Immune IgG significantly decreases, but does not eliminate gp71 expression in all tissues tested. In control animals, ICC reside within a minor subpopulation of cortical, thymic T cells, whereas peripheral (i.e., splenic) ICC are totally devoid of conventional T cell, B cell, and macrophage phenotypic markers. Although thymocytes appear to be a major target of this antibody therapy, T-cell reactivity is not compromised.

Ecotropic virus-producing cells in the bone marrow of the leukemia-prone AKR strain of mice were significantly enriched by fractionation on discontinuous density gradients of Percoll and were found in a low-density population of cells comprised predominantly of medium to large blast cells. The high ecotropic virus-producing low-density bone marrow cell population was also found to be significantly enriched in pluripotent stem cells, prothymocytes, and stromal elements. During the period of time defined by a window for successful leukemosuppressive immunotherapy of AKR mice, virus-producing cells were exclusively detected in this fraction of bone marrow cells, implicating the functional classes of cells coenriched in this fraction as both potential targets of anti-viral immunotherapy and responsible for the seeding of the spleen and thymus with infectious ecotropic virus.

Passive anti-viral immunotherapy greatly suppresses the incidence of spontaneous leukemia in AKR mice, rendering the thymus of successfully treated animals devoid of infectious ecotropic retrovirus. Reconstitution assays have determined that the thymic and splenic homing cells of the AKR bone marrow become ecotropic virus producers subsequent to their seeding of these hematopoietic organs and that in vitro depletion of gp71 expressing bone marrow cells reduces stem cell numbers without affecting prothymocyte content. In the thymus, a population of radioresistant cells, which phenotypically resemble cortical thymocytes, but are unique in their expression of high levels of H-2Kk antigen, have been found to produce high levels of both ecotropic and MCF virus and have been implicated as a putative therapeutic target cell population of anti-viral treatment. In addition, the failure of treated animals to reconstitute following lethal irradiation suggests that an immunotherapy-induced alteration occurs in the bone marrow of AKR mice.

In previous reports in this series, we have demonstrated that treatment of young AKR mice with IgG prepared against the viral envelope glycoprotein suppresses the development of spontaneous leukemia. Moreover, animals exhibiting high anti-viral antibody titers can transfer protection to their offspring. In this report we have extended the studies to another strain of AKR mice and find that the ability to transfer protection to offspring was not obtained. Foster nursing experiments were therefore conducted and their outcomes are indicative that maternal factors may be responsible for this phenomenon.

The bone marrow of AKR mice is the richest source of infectious ecotropic cell centers (ICCs) during the neonatal period. The bone marrow ICCs reside in a low-density population expressing high levels of viral glycoprotein (gp71) and Class I histocompatibility antigens (H-2Kk). In addition, ICCs are enriched in the lymphoid band of Ficoll-Hypaque-fractionated bone marrow, the adherent population of nylon wool separated cells and among the low-density subpopulation of Percoll-fractionated marrow. The observed dichotomy between viral antigen expression and actual virus production suggests that actively cycling cells may be the primary virus producers in the AKR bone marrow. The phenotypic and physical data indicate that bone marrow stem cells and/or prothymocytes may be among the initial virus producing cells in the AKR bone marrow. Leukemosuppressive antiviral immunotherapy delays the appearance of ICCs in the bone marrow but does not exert any major long-term changes on the populations of cells present.