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Biomedical subjects

A S Youmans

Publications and source records attributed to A S Youmans.

At least 19 recordsLinked to original sources
PubMed1978

Effect of metabolic inhibitors on the formation of antibody to sheep erythrocytes, on development of delayed hypersensitivity, and on the immune response to infection with Mycobacterium tuberculosis in mice.

The effect of a number of metabolic inhibitors was determined on: (i) the production of cellular immunity to infection with Mycobacterium tuberculosis in mice by vaccination with mycobacterial ribonucleic acid (RNA), (Ii) the production of cellular immunity to infection with M. tuberculosis in mice with viable H37Ra cells, (iii) the induction of antibody formation to sheep erythrocytes, and (iv) the induction of delayed hypersensitivity in mice to purified protein derivative. The pattern of inhibition produced by metabolic inhibitors on cellular immunity to infection with M. tuberculosis produced by mycobacterial RNA was entirely different from the pattern of inhibition produced by the same metabolic inhibitors on antibody formation to sheep erythrocytes. The effect of the metabolic inhibitors on the induction of delayed hypersensitivity to purified protein derivative did not correlate with the pattern of inhibition produced by the same compounds on antibody formation or on the development of immunity produced by mycobacterial RNA. Cellular immunity to infection produced in mice by viable H37Ra cells was not reduced by any of the metabolic inhibitors except actinomycin D. The possible reasons for the lack of activity of the metabolic inhibitors on the immune response to viable H37Ra cells and the lack of correlation with the pattern of inhibition found in mice vaccinated with mycobacterial RNA is discussed.

Animals↗
PubMed1976

Effect of rifampin on immunity to tuberculosis and on delayed hypersensitivity to purified protein derivative.

Mice vaccinated with mycobacterial ribonucleic acid (RNA) produced a high immune response and did not develop delayed hypersensitivity to purified protein derivative (PPD), and rifampin had no effect on the immune response. Mice vaccinated with viable H37Ra cells produced a high immune response and did develop delayed hypersensitivity to PPD. Rifampin had no effect on this immune response, but reduced the footpad reactions to PPD. Both mycobacterial RNA and poly(A:U) served as adjuvants for induction of hypersensitivity to PPD. This hypersensitivity was reduced by the administration of rifampin. Rifampin had no effect on the production of mycobacterial growth inhibitory factor, which is produced following vaccination of mice with mycobacterial RNA or viable H37Ra cells. Rifampin had no effect on the nonspecific phase of the granulomatous response, but did inhibit the secondary allergic phase of this response. The action, therefore, of rifampin that inhibits the induction of delayed hypersensitivity but had no effect on the immune responses against tuberculosis leads to a separation of tuberculin hypersensitivity from cellular immunity to tuberculosis.

Animals↗
PubMed1976

Mycobacterial ribonucleic acid: comparison with mycobacterial cell wall fractions for regression of murine tumor growth.

Mycobacterial ribonucleic acid (RNA) and cell wall skeleton fraction isolated from H37Ra caused P-815 mastocytoma regression in DBA/2 mice provided the animals were presensitized with freshly harvested living H37Ra cells. In the absence of presensitization, only the RNA fraction inhibited. Cell wall skeleton fraction, under these conditions, stimulated tumor growth. Cell wall lipids (from H37Ra) added to H37Ra cell wall skeleton fraction did not increase the inhibitory activity of cell wall skeleton fraction alone. Mycobacterial RNA appeared to be an effective inhibitor of P-815 mastocytoma metastases as shown by (i) the inhibition of a second footpad lesion distant from the one treated and (ii) increase in survival time.

Animals↗
PubMed1976

Mycobacterial RNA. A comparison with intact mycobacteria for suppression of murine tumor growth.

Our experimental data imply the following: (1) An RNA-rich fraction isolated from H37Ra, and intact H37Ra, are effective agents for suppression of tumor growth. (2) For suppression of two chemically-induced tumors with both RNA and H37Ra, direct contact with tumor is necessary. This may not be the case with other tumors in different strains of syngeneic mice. (3) The host's immunological system is a necessary component for inhibition of tumors by our test materials since there is no tumor suppression in immunologically impaired mice.

Animals↗
PubMed1973

Relationship between tuberculin hypersensitivity and cellular immunity to infection in mice vaccinated with viable attenuated Mycobacterial cells or with Mycobacterial ribonucleic acid preparations.

The migration inhibition technique has been used to study delayed hypersensitivity in vitro by using peritoneal exudate cells and splenic lymphocytes from mice vaccinated with viable cells of the attenuated H37Ra strain of Mycobacterium tuberculosis and from mice vaccinated with ribonucleic acid (myc RNA) preparations obtained from viable mycobacterial cells of the same strain. Inhibition of macrophage migration was noted when purified protein derivative (PPD) or viable H37Ra cells were added to peritoneal exudate cells obtained from mice immunized with viable H37Ra cells and not from mice immunized with myc RNA. Splenic lymphocyte cultures were exposed to the same antigens in vitro. Filtered supernatant fluids from these lymphocyte cultures, when added to peritoneal exudate cells obtained from nonimmunized mice, inhibited migration only when they were obtained from lymphocytes which came from mice immunized with viable H37Ra cells. Injection of PPD intravenously into vaccinated mice resulted in inhibitory supernatant fluids from splenic lymphocyte cultures only when the lymphocytes came from mice immunized with viable H37Ra cells. However, intravenous injection of either viable H37Ra cells or of myc RNA preparations into mice vaccinated with myc RNA occasionally produced inhibitory supernatant fluids when lymphocytes were obtained from these mice. On the other hand, mice vaccinated with myc RNA or viable H37Ra cell preparations were consistently and equally protected against intravenous challenge with the virulent H37Rv strain. Thus, although some evidence was obtained for a delayed type hypersensitivity in mice vaccinated with H37Ra cells or with myc RNA to ribosomal proteins or other proteins associated with the RNA preparation, no evidence of tuberculin hypersensitivity could be detected in any mice vaccinated with the myc RNA. These results argue against a role for tuberculin hypersensitivity in immunity to tuberculous infection.

Animals↗
PubMed1972

Response of vaccinated and nonvaccinated syngeneic C57B1-6 mice to infection with Mycobacterium tuberculosis.

Data are presented which show that the syngeneic C57B1/6 mouse strain is far more susceptible to infection with Mycobacterium tuberculosis strain H37Rv than are the other allogeneic mouse strains used in this laboratory, particularly the Strong A and the CF-1 strains. Not only are the C57B1/6 mice more susceptible to tuberculous infection, but also they respond to infection more uniformly than do the allogeneic strains. C57B1/6 mice develop immunity to challenge with small infecting doses of the virulent H37Rv strain when they are vaccinated with viable cells of the attenuated H37Ra strain and with ribonucleic acid (RNA) preparations isolated from the H37Ra strain. Mice vaccinated with viable cells of the H37Ra strain, however, may die more rapidly than nonvaccinated mice when given a large infecting dose (1.0 mg). This accelerated type of disease is not seen in mice vaccinated with mycobacterial RNA. Since C57B1/6 mice are known to develop tuberculin hypersensitivity more readily than many other mouse strains, the possibility is discussed that the increased susceptibility to tuberculous infection of mice vaccinated with viable cells of the H37Ra strain may be due to a superimposition of a pronounced acute inflammatory response due to tuberculin hypersensitivity upon the infectious process. The several advantages that may be gained in the study of certain host-parasite interactions in tuberculosis by the use of a highly susceptible syngeneic mouse strain such as the C57B1/6 are discussed.

Animals↗
PubMed1972

Effect of polybasic amines on the immunogenicity of mycobacterial ribonucleic acid.

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.

Amines↗
PubMed1971

Failure of synthetic polynucleotides to affect the immunogenicity of mycobacterial ribonucleic Acid and ribosomal protein preparations.

The synthetic polyribonucleotides adenylic acid (poly A), uridylic acid (poly U), cytidylic acid (poly C), and inosinic acid (poly I), whether single- or double-stranded (poly A:U, poly I:C), cannot replace mycobacterial ribonucleic acid (RNA) in the production of a high immune response in CF-1 mice against tuberculous disease. These conclusions are based on the results of several types of experiments. (i) Poly A and poly U, used either singly or in combination, did not increase the immunogenicity of mycobacterial RNA preparations whether emulsified in Freund's incomplete adjuvant (FIA) or not emulsified. (ii) Mycobacterial ribosomal protein, extracted with 2-chloroethanol, was not immunogenic; the addition of poly A:U to the protein did not produce an immune response and FIA did not affect these results. (iii) The RNA left after the protein was extracted was partially immunogenic when emulsified in FIA even though it was partially degraded. (iv) Mycobacterial RNA prepared with ethyl alcohol and partially degraded with ribonuclease had a significantly lower immunogenic activity, and the original higher immune response was not restored by the addition of poly A:U. (v) Mycobacterial RNA totally degraded by weak alkali was not immunogenic, the original immunogenic activity was not restored by the addition of poly A:U or poly I:C, and FIA again did not influence the results. These findings suggest that (i) protein, polypeptides, or other antigenic fragments, if present, are not the specific immunogens; and (ii) mycobacterial RNA is responsible for the high immunogenic activity of mycobacterial ribosomal and RNA preparations. In addition, since the double-stranded forms of these synthetic polynucleotides markedly potentiate the formation of circulating antibodies, these results also reemphasize the lack of correlation between conventional antibody formation and immunity against tuberculosis.

Journal Article↗
PubMed1970

Immunogenic mycobacterial ribosomal and ribonucleic Acid preparations: chemical and physical characteristics.

Five to 20% linear sucrose gradients were used to obtain sedimentation patterns of mycobacterial ribosomes, ribosomal subfractions, and ribonucleic acid (RNA) preparations. Classical 70S ribosomes were obtained when 10(-1)m magnesium chloride was used. These, when dialyzed against 10(-4)m MgCl(2), yielded typical 50S, 30S, and smaller ribosomal subunits. The 30S subunits were the most immunogenic under these conditions. A ribosomal preparation containing subunits which varied from 2.5 to 40S was fractionated by collecting five fractions from a sucrose gradient; based upon the amount of nucleic acid present, the fraction containing the 40S particles was most immunogenic. Physical and chemical evidence suggested that mycobacterial RNA preparations extracted with 65% ethyl alcohol from the ribosomes and diluted in distilled water, were either double-stranded, or mostly double-helical, or had a highly organized secondary structure. This was based on the following observations. (i) Native RNA was resistant to trace amounts of ribonuclease. (ii) The approximate T(m) value in SSC buffer (0.15 m NaCl plus 0.015 m sodium citrate) was greater than 85 C and in 0.1 SSC buffer was 55 C; the RNA diluted in SSC buffer produced a hypochromic effect on cooling at room temperature. (iii) Formaldehyde, in the presence of SSC buffer, decreased the T(m) of the RNA to approximately 55 C, and there was no hypochromic effect on cooling. (iv) Formaldehyde did not increase the wavelength of maximal adsorption of the RNA. (v) The purine/pyrimidine ratio was close to one. (vi) The major peak of the RNA sedimented in the more dense zones of the sucrose gradients. There was a relationship between the sedimentation pattern obtained with the RNA-protein subunits on sucrose gradients and immunogenicity; several examples are given. RNA-protein complexes of approximately 14 to 20S, and occasionally 23S in the major peak, appeared to produce the highest immune response. Smaller RNA-protein complexes such as 6S, which were obtained when the RNA preparation was diluted in certain buffers, were much less immunogenic. This was confirmed by collecting five fractions from sucrose gradients and finding the third fraction (containing RNA-protein complexes approximately 15 to 16S) the most immunogenic. Immunogenic activity was apparently related to the structure of the RNA since it was maximal when the RNA appeared to be either double stranded, double helical, or had a highly organized structure.

Journal Article↗
PubMed1969

Immunizing capacity of viable and killed attenuated mycobacterial cells against experimental tuberculous infection.

The relationship of the dose of vaccine to the immune response was determined in CF-1 mice vaccinated intraperitoneally with viable cells of the attenuated H37Ra strain of Mycobacterium tuberculosis and in mice vaccinated with cells of the same strain killed by autoclaving at 121 C for 15 min. The results showed, in terms of increased resistance to tuberculous infection, that the immune response with both living and killed cells was dependent upon the dose of vaccine, whereas only the living cells were dependent upon the time of challenge after vaccination. The dose response curves show dramatically that viable cells, which do not multiply in vivo, are several hundred times more effective immunizing agents against tuberculous infection than are autoclaved cells. Viable 2-week-old H37Ra cells were far more immunogenic than viable 4-week-old cells. Autoclaved 2-week-old cells, however, were no more immunogenic than autoclaved 4-week-old cells. H37Ra cells killed by boiling (98 C), exposure to 65 C for 30 min, treating with 2% phenol, or by being dried with acetone also lost most of their capacity to immunize mice. The effect of adjuvant on the immune response of mice to tuberculous infection was tested by incorporating both viable and autoclaved cells in Freund's incomplete adjuvant. We found that this vehicle had little or no effect on the immunizing capacity of either viable or heat-killed mycobacterial cells. The relationship of all the findings to the specificity of the immune response to tuberculosis is discussed.

Acetone↗
PubMed1969

Allergenicity of mycobacterial ribosomal and ribonucleic acid preparations in mice and guinea pigs.

Guinea pigs were injected subcutaneously with mycobacterial ribosomal fraction incorporated in Freund's incomplete adjuvant and tested 6 and 12 weeks later by the intradermal injection of 0.5 mug (25 TU) of Purified Protein Derivative. No evidence of delayed-type hypersensitivity could be detected in these animals, although large necrotic reactions were obtained in guinea pigs sensitized with living, attenuated mycobacterial cells. Mice also were vaccinated by the intraperitoneal injection of mycobacterial ribosomal fraction or ribonucleic acid (RNA) and tested for sensitivity to tuberculin at various subsequent times. No evidence of true tuberculin hypersensitivity could be detected at any time, although what appeared to be small Arthus type reactions were seen in mice given the largest vaccinating doses. Attempts to recall tuberculin sensitivity in vaccinated mice by the intravenous injection, 4 weeks after vaccination of living cells, of either the virulent or attenuated mycobacterial strains were unsuccessful. Instead, when the virulent cells were injected, a suppression of footpad reactivity was noted in animals made sensitive to tuberculin by the previous intraperitoneal injection of viable attenuated mycobacterial cells. Both guinea pigs and mice, vaccinated as described above, were also skin tested or footpad tested, respectively, with 2 mug of the ribosomal fraction or RNA used for vaccination. No evidence of true tuberculin hypersensitivity could be obtained; instead, in guinea pig skin very small dermonecrotic areas were noted, and in mice swelling and redness of the footpad occurred to an equal extent in both vaccinated and nonvaccinated mice. The possible role of tuberculin hypersensitivity in acquired immunity to tuberculosis is discussed, and the conclusion is reached that its part, if any, is minor.

Allergens↗
PubMed1969

Factors affecting immunogenic activity of mycobacterial ribosomal and ribonucleic acid preparations.

By following careful procedures, mycobacterial ribosomal fractions and ribonucleic acid (RNA) prepared by ethyl alcohol precipitation were obtained which have immunogenic activities similar to the viable attenuated H37Ra cells of Mycobacterium tuberculosis from which they were obtained. This comparison was based on the amount of ribonucleic acid (RNA) present. These preparations consisted of approximately 63% RNA and 37% protein; no deoxyribonucleic acid or polysaccharide was detected by chemical tests. A high correlation was found between the immunogenic activity of a preparation and the per cent increase in hyperchromicity at 260 nm of a ribonuclease-hydrolyzed portion. Final concentrations of sodium dodecyl sulfate higher than 0.25% when used for the preparation of the ribosomal fractions and RNA resulted in significantly lower immune responses and greater variation between experiments. This was not related to the amount of protein present. The stability of the ribosomal and RNA preparations was tested under a variety of conditions. The need for a good protective adjuvant again was shown since mouse serum readily hydrolyzed the RNA. Equal immunity was obtained after immunization by the intraperitoneal and subcutaneous routes; however, no immune response was obtained when the intravenous route was used. Preliminary results with RNA prepared with phenol showed that it was more easily degraded during preparation. This resulted in a lower immune response than was obtained with the RNA prepared with ethyl alcohol.

Animals↗