Expression and modulation of major histocompatibility antigens on murine primary brain tumor in vitro

Abstract

Lysis of tumor cells by activated cytotoxic lymphocytes requires their recognition of antigens associated with major histocompatibility complex molecules. The authors studied the constitutive expression of Class I and Class II major histocompatibility complex antigens on mouse brain-tumor cells and the capacity of different cytokines and cytokine combinations to alter this expression in vitro. Cells from the murine glioma 26 (GL26), glioma 261 (GL261), and ependymoblastoma A (EpA) cell lines were established in monolayer culture and treated for 48 hours with either alpha interferon, gamma interferon, tumor necrosis factor alpha, tumor necrosis factor alpha plus gamma interferon, or interleukin-2. They were then analyzed by flow cytometry for baseline and cytokine-altered major histocompatibility complex expression. All cell lines had a similar constitutive major histocompatibility complex pattern with low Class I antigen expression and no detectable Class II antigen expression. Alpha interferon substantially induced and up-regulated Class I antigen expression, but had no effect on Class II antigen expression. Gamma interferon also stimulated up-regulation of Class I antigen expression, generally doubling the anti-Class I antigen fluorescence of treated cells. Its effect on Class II antigen expression was more extensive. In the GL26 and GL261 cell lines the expression of Class II antigen determinants increased to 12 x and 14 x control values and as many as 75% of cells that had no detectable constitutive expression of Class II antigen expressed this antigen after priming with gamma interferon. The addition of tumor necrosis factor alpha to gamma interferon further increased Class II antigen expression on EpA tumor cells only. Interleukin-2 and tumor necrosis factor alpha alone had no effect on Class I or Class II antigen expression of any cell lines. It is concluded that Class I and Class II antigen expression in mouse glioma cell lines is induced and enhanced after treatment with certain cytokines in vitro. Use of these cell lines to create in situ primary brain tumors in C57BL/6 mice should provide an excellent animal system to study major histocompatibility complex modulation in brain tumor cells and to examine the potential impact of major histocompatibility complex up-regulation on the response of brain tumors to immunotherapy.

Synergism between GM–CSF and IFNγ: Enhanced immunotherapy in mice with glioma

“…16,17,22 FIGURE 2 -Upregulation of MHC I and MHC II expression on GL-GM and GL-wt cells after IFNg treatment and irradiation. GL-wt and GL-GM were incubated with IFNg (100 ng/ml) for 4 days.…”

“…IFNg is a multifunctional cytokine that recently has been linked to immune surveillance of developing tumors. 16 Some effects induced by IFNg and important for tumor eradication are MHC I and II upregulation on tumor cells, inhibition of tumor cell proliferation 17 and enhanced phagocytic-and antigen presenting capacity of APCs. 16 IFNg is also known to favor a Th1 response, resulting in a tumor specific CD8 1 cytotoxic T-cell (CTL) response.…”

Treatment of intracranial glioma with in situ interferon-gamma and tumor necrosis factor-alpha gene transfer

“…This may be due, in part, to the active expression of immune -suppressing cytokines, 34 which inhibit T-cell and antigen -presenting cell function, 35 as well due to the failure of glioma cells to express adequate levels of MHC on their surface, thereby depriving TIL with signals necessary for recognizing tumor antigens. 23 In addition, glioma cells may induce the local expression of certain proteins, such as FasL, which may directly engage and induce apoptosis in T cells, thereby inhibiting their movement into and through neoplastic tissue. 36 The use of cytokine therapy to address glioma immune privilege has been extensively investigated.…”

“…22 Both IFNg and TNFa modulate cell surface expression of MHC -I and -II, which are key molecules involved in the recognition of antigen by and activation of tumor-specific T cells. 20,23 In intracranial tumor models, IFNg and TNFa have shown therapeutic benefit by promoting both immune 24 and nonimmune 25,26 antitumor activity. With the aim of further investigating the tumoricidal mechanisms and therapeutic uses of IFNg and TNFa, separately and in combination, for glioma, we describe the use of replicationdeficient adenoviral vectors encoding either IFNg ( AdmIFNg ) or TNFa (AdTNFa) for the treatment of a syngeneic murine glioma model.…”

Vascular Apoptosis and Involution in Gliomas Precede Neovascularization: A Novel Concept for Glioma Growth and Angiogenesis

“…GL 261 glioma cells were implanted in the brain of 28 C57BL6 mice (Akbasak et al, 1991). The mice were placed in the Kopf rodent stereotactic apparatus (Tujunga, California).…”