Cytokine Production from Murine CD4 and CD8 Cells After Mannan-MUC1 Immunization

Abstract

Immunotherapy with oxidized mannan-MUC1 fusion protein (M-FP) leads to a T1 immune response characterized by the generation of cytotoxic T lymphocytes (CTL), few antibodies, secretion of interleukin-2 (IL-2), IL-12, and interferon-gamma and tumor protection. Immunotherapy with reduced M-FP or fusion protein (FP) alone leads to a T2 immune response characterized by the generation of MUC1 antibodies, few CTL, IL-4 secretion, and no tumor protection. In these studies, cytokine production from T cells was measured from cultures containing whole spleens. We now report the cytokine secretion patterns from spleen cells separated into CD4+ and CD8+ T cells obtained from mice immunized with either oxidized M-FP, reduced M-FP or FP, or the simultaneous administration of oxidized M-FP and FP. Immunization with oxidized M-FP led to the secretion of T1 cytokines from CD8+ T cells (IL-2, IFN-gamma, and tumor necrosis factor-alpha [TNF-alpha]) and from CD4+ T cells (IL-2 and IFN-gamma). IL-12 production, presumably from activated macrophages, was observed in CD8+ but not CD4+ cultures. Immunization with either reduced M-FP or FP led to the secretion of predominantly T2 cytokines from CD4+ T cells (IL-4 and IL-10) and IL-2 production in both CD4+ and CD8+ T cell cultures. The simultaneous immunization of both oxidized M-FP and FP led to the production of both T1 and T2 cytokines from CD8+ T cells (IL-2, IFN-gamma, and TNF-alpha) and CD4+ cells (IL-2, IFN-gamma, IL-4, and IL-10) and IL-12 production in CD8+ cultures that is, both types of immune responses could occur together. The results demonstrate that the cellular immune response observed in oxidized M-FP-immunized mice is indeed dependent on the T1 cytokine profile secreted by CD8+ T cells, and the simultaneous production of both T1 and T2 cytokines is not cross-inhibitory.

Activation of Antigen-Specific T Cell-Responses by Mannan-Decorated PLGA Nanoparticles

“…The ability of mannosylated formulations to direct the antigens into MHC class I and class II for stimulation of CD8 + and CD4 + T-cell responses, respectively, have been previously described (reviewed in (1)). Differential activation of either CD4 + or CD8 + T-cell responses have been observed when different forms of MN have been utilized (either oxidized or reduced) (28,29). In brief, oxidized MN has shown to facilitate the endosomal escape of mannosylated antigens from the endosome to the cytoplasm with the subsequent activation of CD8 + T-cell responses (30).…”

Interferon-α Therapy following Autologous Peripheral Blood Stem Cell Transplantation for Adult T Cell Leukemia/Lymphoma

“…PBMC were then stained for cell membrane CD3 and CD8 with specific monoclonal antibodies. In addition, cells were stained for cytoplasmic IFN-γ or TNF-α after formalin-fixation with specific monoclonal antibodies by the ordinary method [11]. Anti-CD3 (or CD28) and anti-CD8 were purchased from Immunotech (France).…”

Receptor-Mediated Delivery of Antigens to Dendritic Cells: Anticancer Applications

“…22 MUC1 conjugated to mannan under reducing conditions (where no aldehydes or Schiff bases are present) induces strong T2-type immune responses with a high level of IgG1 antibodies, IL-4 cytokine production, a low CTL precursor frequency, and no protection in mice against a tumor challenge. [23][24][25][26][27][28] However, conjugation of MUC1 to mannan under oxidizing conditions (where aldehydes and Schiff bases are present) yields an immunogen that generates a T1-type response, as indicated by CD8 + CTLs, a low level of IgG2a antibodies, and IL-12 and IFNγ cytokine production. [23][24][25][26][27][28] Both conjugate formulations evidently bind equally to the MR and are taken up into early endosomes.…”