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Detection of keyhole limpet hemocyanin (KLH)-specific immune responses by intracellular cytokine assay in patients vaccinated with idiotype-KLH vaccine

  1. Author:
    Neelapu, S. S.
    Baskar, S.
    Kwak, L. W.

  2. Author Address

    NCI, Expt Transplantat & Immunol Branch, Bldg 567, Room 205, Frederick, MD 21702 USA. NCI, Expt Transplantat & Immunol Branch, Frederick, MD 21702 USA. NCI, ETI Branch, Frederick, MD 21702 USA. SAIC, Frederick, MD USA. Kwak LW NCI, Expt Transplantat & Immunol Branch, Bldg 567, Room 205, Frederick, MD 21702 USA.
    1. Year: 2001
  2. Journal: Journal of Cancer Research and Clinical Oncology
    1. 127
    2. Suppl. 2
    3. Pages: R14-R19
  4. Type of Article: Article
  1. Abstract:

    Keyhole limpet hemocyanin (KLH) has been used as an immune potentiator to enhance antigen-specific responses against haptens and weak antigens including self-antigens. In the present study, we describe the optimization of the intracellular cytokine response to KLH in peripheral blood mononuclear cells (PBMC) of lymphoma and myeloma patients that were vaccinated with tumor-specific immunoglobulin (ld) conjugated to KLH. Addition of anti-CD28 antibody significantly enhanced cytokine-producing CD4(+) T cells. While fresh PBMC showed maximal cytokine response 14 h after antigen stimulation, the frozen PBMC showed maximal cytokine responses by 24 h. Supplementation or the culture medium with fetal bovine serum gave a better signal-to-noise ratio than human AB serum in the intracellular detection of cytokines. The intracellular cytokine responses correlated with the cytokine measurements by enzyme-linked immunosorbent assay (ELISA). Together these results indicate that the intracellular cytokine assay is very helpful to measure antigen-specific immune responses, and in subsequent studies, we have utilized this sensitive technique to detect immune responses against tumor antigens such as idiotype.

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