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The culture requirements for the production of canine I1-2 are reported. Several parameters have been tested, such as concentration of lectin, length of culture period and presence of the additives serum, polyethylene glycol (PEG) and phorbol myristic acetate (PMA). Optimal results have been obtained by stimulation of peripheral blood leukocytes with the lectin PHA (8 micrograms/ml) for 48 h. Techniques for the production of I1-2 containing supernatant free of PHA have been evaluated. Gel filtration chromatography of culture supernatant revealed that canine I1-2 has a molecular weight (m.w.) of approximately 30,000 daltons, similar to the m.w. of I1-2 produced by murine T cells.
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