Effects of the Chinese Traditional Medicine Mao-Bushi-Saishin-To on Therapeutic Efficacy of a New Benzoxazinorifamycin, KRM-1648, againstMycobacterium aviumInfection in Mice

Abstract

The Chinese traditional medicine mao-bushi-saishin-to (MBST), which has anti-inflammatory effects and has been used to treat the common cold and nasal allergy in Japan, was examined for its effects on the therapeutic activity of a new benzoxazinorifamycin, KRM-1648 (KRM), against Mycobacterium avium complex (MAC) infection in mice. In addition, we examined the effects of MBST on the anti-MAC activity of murine peritoneal macrophages (Ms). First, MBST significantly increased the anti-MAC therapeutic activity of KRM when given to mice in combination with KRM, although MBST alone did not exhibit such effects. Second, MBST treatment of Ms significantly enhanced the KRM-mediated killing of MAC bacteria residing in Ms, although MBST alone did not potentiate the M anti-MAC activity. MBST-treated Ms showed decreased levels of reactive nitrogen intermediate (RNI) release, suggesting that RNIs are not decisive in the expression of the anti-MAC activity of such M populations. MBST partially blocked the interleukin-10 (IL-10) production of MAC-infected Ms without affecting their transforming growth factor ␤ (TGF-␤)-producing activity. Reverse transcription-PCR analysis of the lung tissues of MAC-infected mice at weeks 4 and 8 after infection revealed a marked increase in the levels of tumor necrosis factor alpha, gamma interferon (IFN-␥), IL-10, and TGF-␤ mRNAs. KRM treatment of infected mice tended to decrease the levels of the test cytokine mRNAs, except that it increased TGF-␤ mRNA expression at week 4. MBST treatment did not affect the levels of any cytokine mRNAs at week 8, while it down-regulated cytokine mRNA expression at week 4. At week 8, treatment of mice with a combination of KRM and MBST caused a marked decrease in the levels of the test cytokines mRNAs, especially IL-10 and IFN-␥ mRNAs, although such effects were obscure at week 4. These findings suggest that down-regulation of the expression of IL-10 and TGF-␤ is related to the combined therapeutic effects of KRM and MBST against MAC infection.on July 3, 2020 by guest http://aac.asm.org/ Downloaded from a Mice infected i.v. with MAC N-444 (10 7 CFU) were given or not given the indicated agents by gavage either once weekly (KRM) or once daily five times per week (MBST) from day 1 for up to 4 weeks. b There were six mice per regimen. c ND, not determined. d Significantly smaller than the value of untreated control mice (P Ͻ 0.05 by Bonferroni's multiple t test). e Significantly smaller than the value of mice given MBST alone (P Ͻ 0.05 by Bonferroni's multiple t test). f The difference from the values of mice given KRM alone was statistically significant (P Ͻ 0.05) when assessed by the Mann-Whitney test.

Roles of tumour necrosis factor-alpha (TNF-α), transforming growth factor-beta (TGF-β), and IL-10 in the modulation of intercellular adhesion molecule-1 (ICAM-1) expression by macrophages during mycobacterial infection

“…The 10 times higher multiplicity of infection than those for the ICAM‐1 expression assay was employed in order to achieve significant levels of TNF‐ α and TGF‐ β production by MAC‐infected macrophages. At intervals, culture fluid was withdrawn and concentrations of TNF‐ α , IL‐10, and TGF‐ β were measured by ELISA as previously described [19], using rat anti‐mouse TNF‐ α MoAb (PharMingen), mouse anti‐human TGF‐ β MoAb (specific to mouse TGF‐ β ) (Genzyme), and rat anti‐mouse IL‐10 MoAb (Genzyme) as capture antibodies.…”

“…ICAM‐1 mRNA in MAC‐infected macrophages was measured by the reverse transcription‐polymerase chain reaction (RT‐PCR) method as previously described [19]. Macrophages (Method B) were infected with 1 × 10 6 CFU/ml of MAC and cultured in 10 ml of 10% FBS–RPMI 1640 medium in 25‐cm 2 tissue culture flasks at a density of 1·6 × 10 5 cells/flask, at 37°C for up to 14 days.…”

Comparative roles of free fatty acids with reactive nitrogen intermediates and reactive oxygen intermediates in expression of the anti-microbial activity of macrophages againstMycobacterium tuberculosis

“…After washing with 2% FBS±HBSS, the resultant macrophages were cultured for 2 h in the medium containing 1´0 Â 10 6 CFU/ml of M. tuberculosis. After infection with M. tuberculosis, the macrophages were washed with 2% FBS±HBSS and then cultivated in the fresh medium for up to 12 h. At intervals, total RNA was isolated from the cultured macrophages and subjected to RT-PCR as described previously [25]. Briefly, after DNase-I treatment of the RNA sample, the resultant RNA samples were reverse transcribed to the first strand of cDNA using oligo dT primers and Superscript II reverse transcriptase (Gibco, Rockville, MD) according to the recommendations of the manufacturer.…”

Development of new antituberculous drugs based on bacterial virulence factors interfering with host cytokine networks

“…In addition, chemotherapy in combination of antituberculous drugs and immunoadjunctive agents, which allow a mild potentiation of the host's protective immunity without eliciting immunosuppressive cytokines, is appreciably useful [8]. Immunostimulants, such as Chinese traditional medicines and ATP, may be suitable for this purpose [8,115,116].…”