Autologous adjuvant linked fibroblasts induce anti-glioma immunity: implications for development of a glioma vaccine.
OBJECTIVE
Adjuvant-linked vaccines have been shown to induce anti-tumor immunity in patients with a variety of solid tumors. In this study we describe an in vitro model of active immunotherapy using autologous fibroblasts as immunogen. Correlative results from glioma patients immunized with autologous fibroblasts are also described.


METHODS
Peripheral blood lymphocytes (PBLs) from normal subjects were immunized in vitro against autologous skin fibroblasts coupled to the adjuvant muramyl dipeptide. The lymphocytes developed cell-mediated cytotoxicity that was measured with a short-term chromium release assay. Results of in vitro experiments were compared to data derived from glioma patients immunized with subcutaneous injection of an autologous adjuvant-linked fibroblast vaccine. Glioma target cells and fibroblast immunogens were derived from early passage primary tissue culture.


RESULTS
A comparison of autologous vs. homologous immunogen indicated that major histocompatibility complex matching was required at the sensitization stage of immunity (17.2 +/- 3.4% specific lysis vs. 0.4 +/- 3.1%, P < 0.01). Pre-treatment of fibroblast immunogen cells with interferon gamma (IFN-gamma) was found to significantly increase immunity (42.2 +/- 10.0%, P < 0.01), as did IFN-gamma pre-treatment of tumor target cells (35.8 +/- 9.0%, P < 0.01). The positive effect of IFN-gamma was diminished by treatment of cells with IFN-alpha. These in vitro results correlated well with in vivo data derived from glioma patients immunized with an autologous adjuvant-linked fibroblast vaccine. PBLs from patients developed direct cell-mediated cytotoxicity against autologous tumor cells. Lysis of tumor targets after in vivo immunization increased over a three-week interval (from 1.2 +/- 3.0% to 21.0 +/- 3.4%, P < 0.01) while lysis of a non-MHC matched control cell line remained essentially unchanged.


CONCLUSIONS
Specific lysis of glioma targets in vitro was achieved after in vivo sensitization with autologous adjuvant-linked fibroblasts. Collectively, the data indicate that biochemically modified autologous cells can stimulate anti-glioma immunity in humans. The degree of specific immunity seen in our patients compares favorably with other published series using glioma cells as an antigenic source. Accordingly, fibroblasts may represent a practical alternative to glioma cells for vaccine construction.
Parsa
Andrew T
AT
Department of Neurological Surgery, University of California, San Francisco, CA 94143, USA. parsaa@neurosurg.ucsf.edu
Miller
John I
JI
Eggers
Arnold E
AE
Ogden
Alfred T
AT
Anderson
Richard C
RC
Bruce
Jeffrey N
JN
D000119:Acetylmuramyl-Alanyl-Isoglutamine / Q000276:immunology*; D000328:Adult; D019496:Cancer Vaccines / Q000627:therapeutic use*; D005260:Female; D005347:Fibroblasts / Q000187:drug effects / Q000276:immunology*; D005910:Glioma / Q000276:immunology* / Q000628:therapy*; D006650:Histocompatibility Testing; D006801:Humans; D007114:Immunization; D007167:Immunotherapy; D007371:Interferon-gamma / Q000494:pharmacology; D008297:Male; D008875:Middle Aged
D019496:Cancer Vaccines; D000119:Acetylmuramyl-Alanyl-Isoglutamine; D007371:Interferon-gamma