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Presents information on a study which determined the temperature dependence of crystal field energy levels of cerium (Ce) ions in metals caused by the Kondo effect. Properties of Ce intermetallic compounds and alloys; Calculation of neutron quasi-elastic and inelastic scattering spectra; Dynamical susceptibility of the ion.

Transfer ribonucleic acid (tRNA) was isolated from a two-heptose mutant of Salmonella typhimurium LT2 (strain SL1004) and was found to afford 100% mouse protection against challenge with 1000 LD50 of strain LT2. The intraperitoneal minimum effective dose of tRNA was 5μg RNA per mouse and this dose was significantly lower than that of ribosomal RNA for ddY mouse strain. The protective immunity was independent of the presence of antibodies to cell-surface antigens, and was transferred mainly by T cells. The protective moiety of tRNA was sensitive to ribonuclease digestion which resulted in 85% reduction in the mouse survival rate, but was completely resistant to protease digestion. The present study demonstrates that the immunogenic activity of salmonella RNA is present in both ribosomal RNA and tRNA. [ABSTRACT FROM AUTHOR]

Conversion of Salmonella typhimurium to L-forms, both in vitro and in vivo, resulted in the expression of proteins cross-reacting to the mycobacterial 65000 MW heat-shock protein (hsp). Immunization of C3H/HeJ mice with a protective dose of stable L-form S. typhimurium induced γδ T cells in the liver, in accordance with the multiplication of L-form Salmonella in Kupffer cells. The number of γδ T cells decreased after the intracellular growth of L-form Salmonella plateaued. Persistance of the L-forms in Kupffer cells, however, allowed hepatic γδ T cells to increase within 48 hr of infection with virulent S. typhimurium. Thus, the intrahepatic colonization of L-form Salmonella seems to keep γδ T cells on standby, but the emergence of these T cells does not correlate with the expression of L-form hsp. In addition, Kupffer cells colonized by L-forms constitutively synthesized mRNA for interleukin-6 (IL-6) and tumour necrosis factor-α (TNF-α). These results suggest that conversion of S. typhimurium to L-forms in phagocytic cells builds up and maintains acquired resistance, conferred by live-cell vaccines of S. typhimurium, against murine typhoid. [ABSTRACT FROM AUTHOR]

The active component in crude ribosomal fraction (CRF) of Salmonella typhimurium, capable of inducing protective antibody, was partially purified by two series of chromatography (Sephadex 0-150 and DEAF-Sepharose CL6B) after sodium dodecyl sulphate (SDS)-treated CRF was precipitated with ammonium sulphate. The major active component was eluted by 0.4-045 M NaCl from DEAE-Sepharose CL6B, and its molecular weight was 43,000 as determined by SDS-polyacrylamide gel electrophoresis. Immunization with the fraction containing 43,000 component alone did not always confer protection on CFI mice, but its administration together with either the purified transfer RNA (tRNA) or Freund's complete adjuvant (FCA) was much more effective against infection with S. typhimurium. Antibody to the fraction containing 43,000 component was not only free in serum but also associated with peritoneal cells. Macrophages that had been exposed to the antibody had enhanced anti-bacterial activity. Western blot analysis showed that 43,000 component did not react to antiserum to lipopolysaccharide (LPS), but to antiserum to CRF. The antibody elicited by non-O antigenic component and the cell-mediated resistance stimulated by the adjuvant effect of RNA together confer effective protection on CFI mice. [ABSTRACT FROM AUTHOR]

Two outbred mouse strains, ddY and CF1, were tested for their ability to be protected against infection with Salmonella typhimurium by several types of salmonella vaccines. These strains have the same levels of innate susceptibility to S. typhimurium, and also have the same capacity to develop delayed-type hypersensitivity (DTH) to salmonella antigens. Both the crude ribosomal fraction (CRF) and live-cell vaccines conferred acquired resistance on both strains, characterized by greater responses of T cells to salmonella antigens. Mice of the ddY strain were also protected by the purified transfer RNA (tRNA) vaccine, which was free of O antigens, but CF1 mice were not, despite the presence of T-cell reactivity with salmonella antigens. Neither strain was protected by the phenol--water-extracted lipopolysaccharide (LPS). The tRNA-immunized CF1 mice were protected by transfer of antiserum to CRF, but not by transfer of anti-LPS antibody. This antiserum to CRF. however, did not transfer acquired resistance into non-immune mice of either strain. These observations suggest that CF1 mice may require an antibody to another non-O antigen existing in CRF to develop acquired resistance, and that stimulation of the defence system by tRNA may be essential to the development of acquired resistance in CF1 mice. [ABSTRACT FROM AUTHOR]

In order to compare the potential of salmonella vaccines prepared from Salmonella zyphimurium to provide the longer-lasting protection from the aspects of cell-mediated immunity, groups of mice were immunized with optimal doses of the following preparations: live cells, ribosome-rich extract, acetone-killed cells, and heat-killed cells. At various intervals post-immunization, mouse peritoneal macrophages and splenic I cells were tested for biological activities. The capacity of each vaccine to confer mouse protection against a lethal challenge with S. typhimurium correlated with the degree of macrophage activation engendered by each of them in the early stage of immunization. In the late stage of immunization, the level of mouse protection conferred by each vaccine was found to be based on the capacity of T cells to respond to salmonella antigens, which correlated with the degree of adoptive immunity by T cells. The live-cell and ribosomal vaccines were superior to killed-cell vaccines in inducing the cell-mediated protection. Thus, the longer-lasting immunity provided by the live-cell and ribosomal vaccines can be accounted for by the fact that T cells of mice immunized with both vaccines have the persistent reactivity to salmonella antigens. [ABSTRACT FROM AUTHOR]

Intravaginal inoculation of approximately 10(6) piliated gonococci into female mice at different stages of the estrous cycle without any antibiotic pretreatment resulted in gonococcal endometritis. The percentage of mice with positive cultures for Neisseria gonorrhoeae one week after challenge was at least 80%, regardless of at which estrous stage the mice were inoculated. Recovery of gonococci from the uterus continued for more than one month, and the recovery rate appeared to depend on the estrous stage at inoculation. Serum levels of antibodies to crude outer membrane complex began rising one week after inoculation with gonococci and reached a maximum three week after challenge. This animal model is proposed for the study of gonococcal genital infection. [ABSTRACT FROM AUTHOR]

The anti-inflammatory effect of erythromycin was investigated using zymosan-induced peritonitis in mice. When mice were given erythromycin 10 mg/kg/day po for 28 days, a marked suppression of inflammatory responses, including the reduced influx of leucocytes, plasma exudation and prostaglandin E2 synthesis, was observed. However, neither a 7-day treatment with erythromycin nor a 28-day treatment with clindamycin suppressed the response. The anti-inflammatory activity induced after a 28-day treatment with erythromycin was comparable to the anti-inflammatory effect conferred by a 2-day treatment with dexamethasone 40 microgram/mouse/day. Thus, these data confirm previous studies which show that erythromycin can exert an anti-inflammatory effect when used over long periods of time. [ABSTRACT FROM AUTHOR]

The effects of erythromycin stearate over a concentration range of 0.1-10 mg/l on production of elastase, protease and leucocidin by clinical isolates of Pseudomonas aeruginosa were investigated. Growth of P. aeruginosa N42 in broth was not affected significantly during 24 h culture with erythromycin (0.1-10 mg/l), although extracellular protein contents were reduced by erythromycin at concentrations of 0.1-1.0 mg/l. Production of elastase and protease by strain N42 was significantly suppressed by erythromycin with a maximum inhibition at 0.5 mg/l, but the complete inhibition of enzyme production was not achieved. In contrast, leucocidin production by strain N42 was completely impaired by erythromycin at concentrations of 0.1-5.0 mg/l. Although the leucotoxic activity, as determined by vital staining, was not detected, the leucocidin fraction prepared from the autolysate of strain N42 cultured with 10 mg/l of erythromycin induced morphological changes in human leucocytes, resulting in release of elastase. Erythromycin exerted similar effects on other clinical isolates of P. aeruginosa. These findings indicate that erythromycin might have a role in P. aeruginosa infection, although it has no direct antibacterial activity. [ABSTRACT FROM AUTHOR]

Conversion of Salmonella typhimurium to L-forms, both in vitro and in vivo, resulted in the expression of proteins cross-reacting to the mycobacterial 65,000 MW heat-shock protein (hsp). Immunization of C3H/HeJ mice with a protective dose of stable L-form S. typhimurium induced gamma delta T cells in the liver, in accordance with the multiplication of L-form Salmonella in Kupffer cells. The number of gamma delta T cells decreased after the intracellular growth of L-form Salmonella plateaued. Persistance of the L-forms in Kupffer cells, however, allowed hepatic gamma delta T cells to increase within 48 hr of infection with virulent S. typhimurium. Thus, the intrahepatic colonization of L-form Salmonella seems to keep gamma delta T cells on standby, but the emergence of these T cells does not correlate with the expression of L-form hsp. In addition, Kupffer cells colonized by L-forms constitutively synthesized mRNA for interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-alpha). These results suggest that conversion of S. typhimurium to L-forms in phagocytic cells builds up and maintains acquired resistance, conferred by live-cell vaccines of S. typhimurium, against murine typhoid.

The difference in natural resistance to Salmonella typhimurium between S. typhimurium-resistant A/J mice and S. typhimurium-susceptible C57BL/6 mice was analyzed. In both strains, the growth of S. typhimurium was controlled in the spleen until 48 hr of infection, while serum C3b levels were increased in A/J mice immediately after infection but not in C57BL/6 mice. Incubation of A/J mouse serum with S. typhimurium or its lipopolysaccharide (LPS) generated sufficient amounts of C3b, but that of C57BL/6 mouse serum with them did not. A/J macrophages had higher intracellular killing activity in vitro than did C57BL/6 cells against S. typhimurium pre-opsonized with each corresponding fresh serum. However, the cells from both mice exhibited a similar level of killing activity against S. typhimurium pre-opsonized with fresh A/J serum or rabbit complement. The resistance of C57BL/6 mice was significantly increased by opsonizing S. typhimurium with fresh A/J serum or rabbit complement before inoculation. The serum level of interferon-gamma (IFN-gamma) in A/J mice was 2.7 times as high as in C57BL/6 mice at 48 hr post-infection. Recombinant murine IFN-gamma enhanced the intracellular killing activity of macrophages from both mice when S. typhimurium was pre-opsonized with fresh A/J serum but not with fresh C57BL/6 serum. These findings suggest that A/J macrophages exhibit maximal killing activity against A/J serum-opsonized S. typhimurium in vivo when the cells are activated with IFN-gamma. Therefore, the rapid and sufficient activation of complement by Salmonella LPS may render A/J mice more resistant against murine typhoid.

L forms of Salmonella typhimurium LT2 conferred strong protection to a lethal challenge with its parental bacterium on innately hypersusceptible C3H/HeJ mice, and its minimal protective dose was approximately 150 L-forming units. Although L-form S. typhimurium was avirulent for C3H/HeJ mice, it multiplied slowly in both the liver and spleen with the maximal growth 2-3 weeks after immunization and thereafter it persisted in the liver until 24 weeks. Protective immunity began to work between 4 and 6 weeks after immunization, and it remained active as long as the L forms colonized the liver (until 24 weeks after immunization). Vaccination with the L form induced a population of T cells responding to L-form whole-cell lysate (WCL), while delayed-type hypersensitivity (DTH) to the extract of S. typhimurium was induced after the establishment of solid immunity. Moreover, neither T-cell responses nor DTH to heat-killed S. typhimurium was generated. In addition, antibody responses were elicited to WCL but not to heat-killed S. typhimurium. These results indicate that protection conferred by the L forms is attributable to the persistent colonization of the L forms rather than the presence of DTH, and also that Salmonella cytoplasmic antigens are involved in induction of immunological responses by vaccination with the L forms.

The effector cells responsible for protection to Salmonella typhimurium in C3H/HeJ mice, conferred by L-form S. typhimurium, were determined by cell transfer test. Nonfractionated spleen cells from 6-week immune mice but not from 24-week immune animals transferred anti-S. typhimurium immunity. Treatment with anti-macrophage antiserum and complement most effectively abolished protective capacity in 6-week immune cells, while anti-T cell monoclonal antibody plus complement reduced it to a lesser extent. However, adoptive protection was achieved only by transfer of immune macrophages along with Lyt-2+ T cells selected from 6-week immune spleen cells. These Lyt-2+ T cells were cytotoxic to Kupffer cells from C3H/HeJ mice which had been infected 48 hr previously and from the mice which had been immunized 1 week previously, but not to the cells from 6-week immune mice and from normal animals. Moreover, protective capacity in immune macrophages seemed to be correlated to the degree of colonization by the L forms, and the inability to transfer immunity of 24-week immune spleen cells may be due to the decrease in the L form-colonization. These results suggest that cooperation between the L form-colonized macrophages and L form-induced cytotoxic Lyt-2+ T cells contributes to anti-S. typhimurium immunity, and might imply the immunological difference between the 6-week immune phagocytes and the cells at an early stage of infection or immunization.

When endotoxin low-responder C3H/HeJ mice were immunized with L-form Salmonella typhimurium, the mice were more susceptible to a lethal challenge with S. typhimurium 1 week after immunization (1-week mice) than were the unimmunized controls. One-week immune mice produced overwhelming amounts of tumor necrosis factor-alpha (TNF-alpha) in the blood after infection, while 4-week immune mice produced lesser amounts of this cytokine with a 75% survival rate at 60 days postinfection. Pretreatment with anti-TNF-alpha antibody prevented 1-week immune mice from succumbing to acute illness. Endotoxin-stimulated peritoneal macrophages from 1-week immune mice produced higher amounts of TNF-alpha in vitro than did those from 4-week immune mice and they expressed larger amounts of TNF-alpha mRNA on Northern blot. The capacity of macrophages to produce TNF-alpha in vitro was correlated with the degree of colonization by the L form in the cells. These results suggest that the colonization by L-form S. typhimurium in macrophages alters the susceptibility to S. typhimurium of C3H/HeJ mice and that TNF-alpha might play a major role in this alteration of host resistance.

Erythromycin (0.2-20 micrograms/ml) induced the proliferation of macrophages of mouse peritoneal exudate cells (PEC) in a liquid medium without exogenous growth factors. The proliferating macrophages formed giant colonies between days 22 and 26 of culture; these colonies continued to proliferate even after subculture. The erythromycin-induced cell proliferation was independent of fibroblasts, T cells, B cells, or endotoxins. This activity seemed to be specific to erythromycin since other antibiotics such as tetracycline, streptomycin, gentamicin, penicillin G, and josamycin did not induce the proliferation of macrophages. Any known cytokines, including IL-2, IL-3, IL-4, IL-6, and GM-CSF, were not detectable by ELISA tests in any of the culture supernatants sampled from day 7 through day 28. The culture supernatants, however, had the capability of inducing the growth of macrophages, only in the presence of bioactive erythromycin at concentrations higher than 1.6 micrograms/l. Moreover, the culture supernatants, sampled after giant colonies had been formed, were capable of inducing giant colonies in the culture of adherent PEC. Thus, the erythromycin-induced macrophage proliferation might be due to the direct effect of this antibiotic, whereas the formation of giant colonies might be due to the production of some unidentified soluble factor produced by the proliferating macrophages. These data indicate that mouse PEC contain a subset of peritoneal macrophages capable of responding to erythromycin by forming proliferating colonies without exogenous growth factors.

We studied the effect of erythromycin (EM) on the attachment of Pseudomonas aeruginosa and Neisseria gonorrhoeae to HeLa and HT-177 cell and on cytotoxin production of P. aeruginosa. 1. EM inhibited attachment of these bacteria. 2. EM inhibited manifestation of the pili of these bacteria. 3. EM inhibited production of protein II, the second attachment factor of N. gonorrhoeae. 4. EM inhibited production of 66 K cytotoxin of P. aeruginosa. On the basis of these findings, it was suggested that EM might inhibit infection by repressing manifestation of the attachment factor and production of cytotoxin of the bacteria.

A cytotoxic protein was isolated from the sodium dodecyl sulphate (SDS)-solubilized extract of the stable L forms of Salmonella typhimurium by ion-retardation chromatography, ion-exchange chromatography, isoelectric focusing and gel filtration. The purified toxin, with a molecular mass of 32 kDa and with isoelectric point of 6.4, was thermolabile and trypsin-sensitive. Against mouse macrophages, its cytolytic effect was detectable in vitro at concentrations higher than 0.7 micrograms/ml, with a complete lysis obtained at 5 micrograms/ml. In contrast, it stimulated C3H/HeJ macrophages in the dose range of 0.1-0.5 micrograms/ml to allow the cell to respond to endotoxin, resulting in the significant production of tumor necrosis factor alpha. By Northern blot analysis, this effect was detectable at a dose as low as 0.01 micrograms/ml. These findings suggest that the transformation of bacillary S. typhimurium into L forms in vivo may induce alterations in host resistance against murine typhoid.

The therapeutic effect of granulocyte colony-stimulating factor (G-CSF) against intramuscular infection with Pseudomonas aeruginosa in cyclophosphamide (CY)-treated mice was analyzed by measuring plasma levels of amyloid P-component (APC) and proinflammatory cytokine levels. CY (100 mg/kg) treatment of mice significantly suppressed plasma concentrations of APC and tumor-necrosis factor-alpha (TNF-alpha) following infection with P. aeruginosa, in associated with enhanced susceptibility of the treated mice to this bacterium. A 4-day treatment of CY-treated mice with recombinant human G-CSF (rhG-CSF) increased resistance of CY-treated mice, together with the marked restoration of APC and TNF-alpha productions. The capacity to produce interleukin 1-beta and TNF-alpha of peritoneal macrophages and also that to produce IL-6 of spleen cells were significantly enhanced by the in vivo administration of rhG-CSF in CY-treated mice. These results indicate that G-CSF may increase the functions of monocytes/macrophages directly or indirectly in vivo. Therefore, the therapeutic effect of rhG-CSF seems to consist of not only increases in the number and functions of neutrophils but also enhancement of monocyte/macrophage functions.

Live-cell vaccines of Salmonella typhimurium, either a sub-lethal dose of a wild-type (strain LT2) or a high dose of its two-heptose Rd1 mutant (strain SL1004), induced acquired resistance to murine typhoid, which remained 180 days after immunization. Growth of S. typhimurium as a bacillary form ceased between days 30 and 60 of immunization, but L forms of this bacterium colonized the liver (the mean number of L forms in the liver: 600 L-forming units) even at 180 days post-immunization. In contrast, a high inoculum of either a Ra mutant (strain TV148) of strain LT2 or S. schottmülleri 8006 sharing the same O antigenic components with those of S. typhimurium induced only a short-lived protection in proportion to the number of L forms in the liver, and the protective immunity was lost before day 180. However, there was no significant difference in the salmonella-specific T-cell responses among groups of immunized mice on day 180 of immunization. A lethal infection with strain LT2 in mice which had been immunized 75 days previously with living cells of strain SL1004 resulted in a rapid clearance of the challenge inoculum, together with a rapid elevation of anti-S. typhimurium antibody responses. Thus, the present data suggest that the long-lived immunity conferred upon live S. typhimurium vaccines is attributable to the colonization of this bacterium in the liver as L forms and the ability to colonize the liver as L forms is independent of the chain length of salmonella O-antigens.

Resistance of mice to Salmonella typhimurium in the early phase of infection is known to be controlled by the expression of chromosome 1 locus Ity. To clarify the mechanism by which the genetically resistant (Ityr) mice can overcome the first phase of salmonellosis, the early response in DBA/2 (Ityr) and BALB/c (Itys) mice was compared after a subcutaneous injection of S. typhimurium. In both strains, the growth of S. typhimurium was controlled in livers and Kupffer cells until day 3, but thereafter the bacteria multiplied rapidly in BALB/c mice. Over the first 2 days nonspecific responses (changes in levels of blood leukocytes, plasma iron, and alpha 1-antitrypsin) were not significantly different between the strains, and the capacity of Kupffer cells isolated from infected mice of both strains to produce interleukin 1 (IL-1) and tumor necrosis factor alpha (TNF-alpha) was of the same degree. Thereafter, only DBA/2 Kupffer cells were able to produce membrane-associated IL-1 (ma IL-1) as well as TNF-alpha. Moreover, only DBA/2 splenocytes were able to produce interferon gamma (IFN-gamma) upon stimulation with Salmonella antigens, although concanavalin A-stimulated splenocytes of both strains produced the same level of interleukin 2. Furthermore, administration of recombinant murine IFN-gamma and DBA/2 Kupffer cells of day 6 to BALB/c mice 3 days after infection resulted in a significant level of protection, whereas neither of these materials alone induced protection. Injection of anti-TNF-alpha antibodies did not affect the resistance of DBA/2 mice. Thus, these findings suggest that the early resistance of Ityr mice is partly attributable to their capacity to produce IFN-gamma and ma IL-1 after infection.

The present study was carried out to ascertain whether the active component of Corynebacterium kutscheri (CK-M) could stimulate host cells of mice to produce several cytokines. CK-M stimulated thioglycollate-induced peritoneal macrophages to produce interleukin 1 (IL-1) and tumor necrosis factor alpha (TNF-alpha) at concentrations of 1-100 ng/ml, and it also induced IL-2 and interferon-gamma (IFN-gamma) as well as IL-6 production by splenocytes. Maximum production of each cytokine induced by CK-M was obtained at the following doses: IL-1 at 5 ng/ml, TNF-alpha at 50 ng/ml, IL-2 at 1 microgram/ml, IL-6 at 500 ng/ml and IFN-gamma at 750 ng/ml. In contrast, IL-4 was not produced to a significant extent by CK-M-stimulated splenocytes. Furthermore, when mice were intravenously injected with 20 micrograms of CK-M, IL-2 and IFN-gamma production by splenocytes, upon stimulation with either formalin-killed C. kutscheri or mitogens, was significantly higher on day 10 of treatment than on day 2. Additionally, the cytotoxicity to L929 cells of this serum from CK-M-treated mice increased with time, and the activity in the serum of day 10 was not abrogated by the antibody to TNF-alpha. Data obtained here indicate that CK-M may preferentially stimulate type-1 helper T cells to produce IL-2 and IFN-gamma, and that the enhanced cytokine production could contribute to the nonspecific resistance induced by C. kutscheri.

The 43-kDa non-O antigenic component isolated from the crude ribosomal fraction of Salmonella typhimurium [9] was further purified by affinity chromatography (43-kDa protein: 43-kDp). Immunization with 43-kDp did not induce complete mouse protection in CF1 mice to 500 LD50 of S. typhimurium, although it elicited a substantial IgG antibody response. The 43-kDp exhibited the mitogenicity to splenocytes (CF1 and C3H/HeJ) and B cell-rich populations (CF1). Complexing 43-kDp with the compact ribosomes of Streptococcus pyogenes by formaldehyde (complex vaccine: CV) elicited both IgM and IgG antibodies to 43-kDp. CV induced a boosting effect to enhance IgG antibody response. Moreover, CV generated delayed-type hypersensitivity to salmonella antigens and also conferred complete protection against 500 LD50 challenge of S. typhimurium to CF1 mice. These abilities of CV were reduced or impaired by RNase digestion. CV was able to induce partial or complete protection in inbred mouse strains (C3H/HeN, C3H/HeJ, DBA/2 and A/J). These data, in addition to other reports, suggest that conformational stability between ribosomes and contaminating substances such as 43-kDp or O-antigens might be required for the overall effects of the ribosomal vaccine.

Clumping and fibrinogen-binding activities of 4 Staphylococcus aureus strains (Cowan I, Newman D2C, Wood 46 and NCTC 5655) were assayed with a semiquantitative clumping test and an enzyme-linked immunosorbent assay (ELISA), respectively. Distinct positive clumping was detected with whole cells of the 3 strains except Wood 46. Amounts of fibrinogen required for a definite clumping depended greatly on strains as well as on their growth phases. On the other hand, fibrinogen-binding activities were detected both in culture supernatants and in cell lysates of all the 4 strains, and the levels were rather comparable with one another and relatively steady through their growth cycles. No significant correlation was thus found among expression behavior of clumping and fibrinogen-binding activities.

This study was conducted to investigate the effects of sex hormones upon the nature of soluble immune response suppressor (SIRS) produced by concanavalin A-stimulated Lyt-2+ T cells. Conventional SIRS affected IgM PFC only. However, SIRS made with progesterone (20-400 ng/ml or Prog-SIRS) suppressed IgM PFC, one-way MLR, and generation CTL; and SIRS made with estrogen (0.2-50 ng/ml or Est-SIRS) enhanced these responses. The factor(s) (MW 40,000-55,000) to stimulate macrophages to produce the second soluble factor (M phi-SF) was isolated from all preparations by gel filtration. Furthermore, Est-SIRS contained a factor(s) (MW 10,000-30,000) to enhance IgM PFC, MLR, and mitogen-induced blastogenesis of both T and B cells; and Prog-SIRS possessed the suppressive factor(s) to IgM PFC, MLR, and mitogen-induced T-cell proliferation. These activities were not impaired by 2-mercaptoethanol. Moreover, the suppressive activity of Prog-SIRS was completely absorbed by T cells only, but the enhancing activity of Est-SIRS was not completely absorbed by a single-cell population. These data suggest that progesterone can contribute to the suppression of allograft rejection through soluble factors, and estrogen can enhance host responses which may be affected by several soluble factors during pregnancy.

The effects of erythromycin stearate (10 mg/kg/day) were studied on productions of interleukin (IL)-1 and -2 in mice after a long-term treatment. A 28-day treatment resulted in higher levels of IL-1 production by macrophages and of IL-2 production by splenocytes, while a 7-day treatment did not increase them. T-cell growth factor activity of IL-2 preparation prepared on day 28 of treatment as determined by HT-2 cell proliferation was reduced by about 40% in the presence of anti-murine IL-4 monoclonal antibodies, while control IL-2 activity was not reduced. Furthermore, a 28-day treatment with erythromycin stearate increased concanavalin A-induced blastogenesis of splenocytes significantly. These results suggest that long-term treatment with erythromycin stearate can stimulate host defense by increasing interleukin production.

The isolation and determination of biological activities of the active component of Corynebacterium kutscheri were attempted in the present investigation. The antitumor effect was confined to the subcellular particle fraction of this bacterium and was associated with a molecule of glycoprotein nature (40,000-38,000 Daltons) isolated from this fraction by affinity chromatography with concanavalin A-Sepharose 4B. This substance exerted mitogenic activity on C3H/HeJ splenocytes and T cells, stimulatory activity on macrophages, and further exhibited antitumor effect on P388 leukemia in CDF1 mice. The Winn assay disclosed that the antitumor effect induced by this substance was dependent on L3T4+ T cells. Furthermore, both the mitogenic and antitumor activity of this moiety were resistant to heating at 100 degrees C for 30 min or RNase digestion, but sensitive to trypsin digestion, or low or high pH. These results indicate that the antitumor effect of C. kutscheri is attributable to the heat-stable glycoprotein moiety which can directly stimulate T cells and macrophages.

Dialyzable factor (DF) prepared from a ribosomal fraction of Salmonella typhimurium was tested for its ability to induce interleukin 1 (IL 1) and 2 (IL 2) production, in relation to acquired resistance, after an intraperitoneal injection of DF. IL 1 production in vitro by peritoneal macrophages of DF-treated mice reached the maximum 4 days after injection, at the time when the nonspecific local resistance via macrophages directly activated with DF became apparent (Kita et al, Microbiol. Immunol. 28:807, 1984). Concanavalin A-induced IL 2 production by splenocytes of DF-treated mice reached the maximal level between days 6 and 8, and it could be enhanced even on day 14. Antigen-induced blastogenic responses of splenocytes from DF-treated mice reached the maximal level 14 days after treatment. Although DF did not show the mitogenic activity to normal splenocytes, T cells of DF-treated mice could respond to S. typhimurium. On the contrary, T cells of normal mice could respond to heat-killed cells of S. typhimurium when they were cultured with macrophages which had been directly stimulated in vitro with DF. Furthermore, T cells from DF-treated mice could respond to antigens of different species of bacteria, and especially to Listeria monocytogenes. These results suggest that T cells of DF-treated mice, being at the intermediate stage of activation via monokines including IL 1 which is produced by macrophages stimulated with DF, are able to proliferate immediately after the administration of challenging organisms as a second signal, and also that the specificity of the response may be defined by the challenging organisms.

Transfer ribonucleic acid (tRNA) was isolated from a two-heptose mutant of Salmonella typhimurium LT2 (strain SL1004) and was found to afford 100% mouse protection against challenge with 1000 LD50 of strain LT2. The intraperitoneal minimum effective dose of tRNA was 5 micrograms RNA per mouse and this dose was significantly lower than that of ribosomal RNA for ddY mouse strain. The protective immunity was independent of the presence of antibodies to cell-surface antigens, and was transferred mainly by T cells. The protective moiety of tRNA was sensitive to ribonuclease digestion which resulted in 85% reduction in the mouse survival rate, but was completely resistant to protease digestion. The present study demonstrates that the immunogenic activity of salmonella RNA is present in both ribosomal RNA and tRNA.