Factors affecting the immunizing activity of ribosomal fractions isolated from Mycobacterium tuberculosis var. bovis strain BCG

Abstract

Levels of antituberculous immunity similar to those induced by live BCG vaccine were detected in CF1 mice immunized with ribosomal fractions isolated from Mycobacterium tuberculosis var. bovis, strain BCG, and challenged 3 weeks later with the virulent H37Rv strain of Mycobacterium tuberculosis var. hominis. The activity of the crude ribosomal preparations was found to be a function of the immunizing doses and the immunity induced by 1.0-mg doses remained at the same high level after 4 weeks of storage at 4 degrees C but decreased markedly thereafter. Dialysis and lyophilization had no detrimental effects on the activity of the crude preparations whereas purification by column chromatography on Sephadex G-200 annihilated their biological activity. Crude low-polysaccharide-containing preparations were found inactive even at the 1.0-mg dose level and results of experiments performed with crude ribosomal fractions of varying polysaccharide contents strongly suggest that polysaccharides, or RNA-polysaccharide complexes, may play an important role in the induction of immunity with crude ribosomal fractions isolated from the BCG strain of Mycobacterium tuberculosis var. bovis.

Biochemical characterization of three mycobacterial ribosomal fractions

“…The carbohydrate-containing substances of ~ibosimal' fractions were injected in incomplete Freund's the H37Ra ribosomal fractions were purified by affinity adjuvant, all emulsions being prepared as previously described chromatography on concanavalin A -Sepharose 4B, and (Portelance et al 1975). Experimental animals were injected the formation of complexes between these substances i.p.…”

“…These suspensions were samples were diluted at 4 mg/mL in the sample buffer used to prepare the ribosomal fractions as previously described (Laernrnli 1970) and heated in a boiling water bath for 2 min. (Portelance et al 1975).…”

“…For the Salmonella typhimurium ribosomal Eisenstein 1975;Feit and Tewari 1974;Fontagnes et al vaccine, the protective activity has been successively 1977; Guentzel and Berry 1974;Hoops et al 1976; ascribed to a ribosomal protein (Johnson 1972(Johnson , 1973, Houchens and Wright 1973;Jensen etal. 1972;Johnson and RNA (Venneman and Berry 1971;1972,1973;Lieberman 1977;Lin and Berry 1978;Lynn Bigley 1969;Vememan et al 1970), an RNA-protein et al 1977;Michel et al 1978;Misfeldt and Johnson complex (Smith and Bigley 1972), a glycoprotein or a 1977; Portelance et al 1975;Portelance et al 1977; mucopolysaccharide (Houchens and Wright 1973), and Riottot et al 1979;Schalla and Johnson 1975; Smith to the 0 antigen contaminating either the crude and Bigley 1972; Swendsen and Johnson 1976; Tewari ribosomes or the RNA (Eisenstein 1975;Hoops et al et al 1977;Tewari et al 1978; Thomas and Weiss 1976;Lin and Berry 1978). Youmans and Youmans (1970) implied that a double-stranded RNA is the isolated from M. tuberculosis.…”

Immunoprotective activity of capsular polysaccharide in Klebsiella pneumoniae ribosomal preparations does not involve ribonucleic acid

“…Since Youmans and Youmans (29) reported that the isolated active fraction of M. tuberculosis is a double-stranded RNA, no one has reported similar results on mycobacteria. Portelance et al (19,20) suggested that polysaccharide or RNA-polysaccharide complexes may play an important role in the induction of immunity against tuberculosis. However, these investigators did not identify the hypothetic polysaccharide involved.…”

Immunity to tuberculosis from the perspective of pathogenesis

“…It is of interest that in spite of the evidence that BCG vaccination protects by reducing bacillemia (13,17), data indicating that experimental vaccines are protective has generally been obtained with animal models in which the animals were challenged intravenously or intraperitoneally, routes which prevent the investigator from measuring the extent to which vaccination inhibits bacillemia (4, 26,51).…”