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Effect of Polybasic Amines on the Immunogenicity of Mycobacterial Ribonucleic Acid

¹ Department of Microbiology, Northwestern University Medical School, Chicago, Illinois 60611

ABSTRACT

The five polybasic amines, methylated bovine serum albumin (MBSA), protamine sulfate, neomycin sulfate, streptomycin sulfate, and diethylaminoethyl-dextran (DEAE-dextran), were examined to determine their effects on the immunogenic activity of mycobacterial ribonucleic acid (RNA) preparations, and whether they could substitute for Freund incomplete adjuvant (FIA) by protecting the mycobacterial RNA in vivo. These compounds were either emulsified in FIA or not emulsified in FIA. Different results were obtained when these compounds, complexed with mycobacterial RNA, were emulsified in FIA. MBSA, in ratios of mycobacterial RNA-MBSA of 1:0.2 to 1:0.4, had no effect on immunogenic activity. However, when the ratio was increased to 1:1, 1:2, or to 1:4, marked inhibition of the immunogenic activity occurred. Protamine sulfate and neomycin sulfate also inhibited the immunogenic activity of mycobacterial RNA; however, neither streptomycin sulfate nor DEAE-dextran had any effect on immunogenic activity. Without being emulsified in FIA, these five polybasic amines, with the exception of DEAE-dextran, acted only as weak adjuvants for mycobacterial RNA and, therefore, could not be used as substitutes for FIA for the protection of mycobacterial RNA from host nucleases. DEAE-dextran, although not as effective as FIA, afforded some protection for the mycobacterial RNA. DEAE-dextran alone also produced a low degree of nonspecific immunity against tuberculosis. Since MBSA, protamine sulfate, and neomycin sulfate reduce the biological activity of mycobacterial RNA after complexing with it, it is probable that these compounds "mask" the immunogenic sites on the mycobacterial RNA structure.