Generation of human class I major histocompatibility complex activating factor in serum free medium and its partial characterization |
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Authors: | Aparna Raval Niti Ipuri R K Saxena |
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Institution: | (1) Immunology Laboratory, School of Life Sciences, Jawaharlal Nehru University, 110067 New Delhi |
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Abstract: | Human peripheral blood mononuclear cells (PBMCs) activated with Con-A release a soluble factor which augments the expression
of class I major histocompatibility complex (MHC) antigens by a variety of tumour cells. Previous attempts to purify this
factor called MHC-activating factor (AF) (MHC-AF) made us realize that the presence of large numbers and quantities of irrelevant
fetal calf serum proteins in the culture supernatants of the activated human PBMCs, interfered with the purification procedure.
It was therefore necessary to standardize the use of a serum free culture medium to generate human MHC-AF. In the present
communication we have tried several types of culture media and have identified DCCM-2 as the most suitable culture medium
to generate human MHC-AF. MHC-AF generated in DCCM-2 medium appears to be a protein molecule resistant to pH 2 treatment but
sensitive to heat treatment (56°C × 45 min) and treatment with proteolytic enzymes trypsin and chymotrypsin. |
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Keywords: | MHC antigens cytokine serum free medium MHC-AF |
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