Abstract
Mycobacterium vaccae is of major pharmaceutical interest as an immunotherapeutic agent. Although M. vaccae 15483 ATCCT strain displays smooth and rough colonial morphologies on solid culture media, it is not known in which conditions M. vaccae switches from one colonial morphotype to the other or whether there are biological differences, especially immunological, between them. We have found that the change from a smooth to rough stable variant occurs spontaneously at 30 °C. The analysis of the composition of the cell wall in both variants showed that the smooth morphotype presents an extracellular material that has never previously been described and was identified as a long-chain saturated polyester that, interestingly, is not produced by the rough morphotype. Our results also indicate that this compound could be implicated in the spreading ability of smooth colonies. Proliferation, IFN-\(\upgamma\) and IL-12(p40) production by splenocyte cultures was significantly higher in mice immunised with the rough variant compared with those immunised with the smooth one. This latter finding suggests that the different colonial morphology of M. vaccae may affect the immunomodulatory effects induced from M. vaccae preparations.
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Abbreviations
- ELISA:
-
enzyme-linked immunosorbent assay
- GC-MS:
-
gas chromatography-mass spectrometry
- HKR:
-
heat-killed rough
- HKS:
-
heat-killed smooth
- HPLC:
-
high performance liquid chromatography
- IR:
-
infrared
- LEDBP:
-
bipolar-gradient LED
- NMR:
-
nuclear magnetic resonance
- PFGSE:
-
pulsed field-gradient spin-echo
- RC:
-
red compound
- SEM:
-
scanning electron microscopy
- TLC:
-
thin layer chromatography
- TTA:
-
2,3,5-triphenyl tetrazole
References
Abbot N.C., Beck J.S., Feval F., Weiss F., Mobayen M.H., Ghazi-Saidi K., Dowlati Y., Velayati A.A., Stanford J.L. (2002) Immunotherapy with Mycobacterium vaccae and pe ripheral blood flow in long-treated leprosy patients, a randomised, placebo-controlled trial. Eur. J. Vasc. Endovasc. Surg. 24: 202–208
Abou-Zeid C., Gares M.P., Inwald J.,Janssen R., Zhang Y., Young D.B., Hetzel C., Lamb J.R., Baldwin S.L., Orme I.M., Yeremeev V., Nikonenko B.V., Apt A.S. (1997) Induction of a type 1 immune response to a recombinant antigen from Mycobacterium tuberculosis expressed in Mycobacterium vaccae. Infect. Immun. 65: 1856–1862
Arkwright P.D., David T.J. (2001) Intradermal administration of a killed Mycobacterium vaccae suspension (SRL 172) is associated with improvement in atopic dermatitis in children with moderate-to-severe disease. J. Allergy Clin. Immunol. 107: 531–534
Balagon M.V., Walsh D.S., Tan P.L., Cellona R.V., Abalos R.M., Tan E.V., Fajardo T.T., Watson J.D., Walsh G.P. (2000) Improvement in psoriasis after intradermal administration of heat-killed Mycobacterium vaccae. Int. J. Dermatol. 39: 51–58
Balagon M.V., Tan P.L., Prestidge R., Cellona R.V., Abalos R.M., Tan E.V., Walsh G.P., Watson J.D., Walsh D.S. (2001) Improvement in psoriasis after intradermal administration of delipidated, deglycolipidated Mycobacterium vaccae (PVAC): results of an open-label trial. Clin. Exp. Dermatol. 26: 233–241
Barrow W.W., Brennan P.J. (1982) Isolation in high frequency of rough variants of Mycobacterium intracellulare lacking C-mycoside glycopeptidolipid antigens. J. Bacteriol. 150:381–384
Belisle J.T., Brennan P.J. (1989) Chemical basis of rough and smooth variation in mycobacteria. J. Bacteriol. 171: 3465–3470
Boenickse R., Juhasz E. (1964) Boeschreibung der neuen Species Mycobacterium vaccae n. sp. Zbl. Bakt. Abt. I, Orig A. 192: 133–135
Camporota L., Corkhill A., Long H., Lordan J., Stanciu L., Tuckwell N., Cross A., Stanford J.L., Rook G.A., Holgate S.T., Djukanovic R. (2003) The effects of Mycobacterium vaccae on allergen-induced airway responses in atopic asthma. Eur. Respir. J. 21:287–293
Cermak S.C., Isbell T.A. (2003) Synthesis and physical properties of estolide-based functional fluids. Ind. Crops Products. 18: 183–196
Chadwick M.V. (1981) Mycobacteria – (Monographs in Medical laboratory science series). Wright-PSG, London
Chambaz E.M., Horning E.C. (1969) Conversion of steroids to trimethylsilyl derivatives for gas phase analytical studies: reactions of silylating reagents. Anal. Biochem. 30:7–24
da Silva T.R., de Freitas J.R., Silva Q.C., Figueira C.P., Roxo E., Leao S.C., de Freitas L.A., Veras P.S. (2002) Virulent Mycobacterium fortuitum restricts NO production by a gamma interferon-activated J774 cell line and phagosome-lysosome fusion. Infect. Immun. 70: 5628–5634
Daffé M., Draper P. (1998) The envelope layers of mycobacteria with reference to their pathogenicity. Adv. Microb. Physiol. 39:131–203
Etienne G., Villeneuve C., Billman-Jacobe H., Astarie-Dequeker C., Dupont M.A., Daffé M. (2002) The impact of the absence of glycopeptidolipids on the ultrastructure, cell surface and cell wall properties, and phagocytosis of Mycobacterium smegmatis. Microbiology 148: 3089–3100
Friebolin H. (1991) Basic One- and Two-dimensional NMR Spectroscopy. VCH, Weinheim
Hadley E.A., Smillie F.I., Turner M.A., Custovic A., Wookcock A., Arkwright P.D. (2005) Effect of Mycobacterium vaccae on cytokine responses in children with atopic dermatitis. Clin. Exp. Immunol. 140: 101–108
Hölscher C. (2004) The power of combinatorial immunology: IL-12 and IL-12 related dimeric cytokines in infectious diseases. Med. Microbiol. Immunol. 193: 1–17
Hrouda D., Baban B., Dunsmuir W.D., Kirby R.S., Dalgleish A.G. (1998) Immunotherapy of advanced prostate cancer: a phase I/II trial using Mycobacterium vaccae (SRL172). Br. J. Urol. 82: 568–573
Janssen R., Kruisselbrink A., Hoogteijling L., Lamb J.R., Young D.B., Thole J.E. (2001) Analysis of recombinant mycobacteria as T helper type 1 vaccines in an allergy challenge model. Immunology 102: 441–449
Knothe G., Nelsen T.C. (1998) Evaluation of the 13C NMR signals of saturated carbons in some long-chain compounds. J. Chem. Soc. Perkin Trans. 2. 9:2019–2026
Luquin M., Ausina V., Lopez-Calahorra F., Belda F., Garcia-Barceló M., Celma C., Prats G. (1991) Evaluation of practical chromatographic procedures for identification of clinical isolates of mycobacteria. J. Clin. Microbiol. 29:120–130
Martinez A., Torello S., Kolter R. (1999) Sliding motility in mycobacteria. J. Bacteriol. 181:7331–7338
Mayo R.E., Stanford J.L. (2000) Double-blind placebo-controlled trial of Mycobacterium vaccae immunotherapy for tuberculosis in KwaZulu, South Africa, 1991–1997. Trans. R. Soc. Trop. Med. Hyg. 94:563–568
Mendes R., O’Brien M.E.R., Mitra A., Norton A., Gregory R.K., Padhani A.R., Bromelow K.V., Winkley A.R., Ashley S., Smith I.E., Souberbielle B.E. (2002) Clinical and immunological assessment of Mycobacterium vaccae (SRL172) with chemotherapy in patients with malignant mesothelioma. Br. J. Cancer 86: 336–341
Moehring J.M., Solotorovsky M.R. (1965) Relationship of colonial morphology to virulence for chickens of Mycobacterium avium and the nonphotochromogens. Am. Rev. Respir. Dis. 92: 704–713
Muñoz M., Raynaud C., Lanéelle M.A., Julián E., Lopez Marín L.M., Silve G., Ausina V., Daffé M., Luquin M. (1998) Seroreactive species-specific lipooligosaccharides of Mycobacterium mucogenicum sp. nov. (formerly Mycobacterium chelonae-like organisms): identification and chemical characterization. Microbiology 144: 137–148
O’Brien M.E., Saini A., Smith I.E., Webb A., Gregory K., Mendes R., Ryan C., Priest K., Bromelow K.V., Palmer R.D., Tuckwell N., Kennard D.A., Souberbielle B.E. (2000) A randomized phase II study of SRL172 (Mycobacterium vaccae) combined with chemotherapy in patients with advanced inoperable non-small-cell lung cancer and mesothelioma. Br. J. Cancer 83: 853–857
O’Brien M.E., Anderson H., Kaukel E., O’Byrne K., Pawlicki M., Von Pawel J., Reck M., SR-ON-12 Study Group. (2004) SRL172 (killed Mycobacterium vaccae) in addition to standard chemotherapy improves quality of life without affecting survival, in patients with advanced non-small-cell lung cancer: phase III results. Ann. Oncol. 15: 906–914
Peláez M., Orellana C., Marques A., Busquets M., Guerrero A., Manresa A. (2003) Natural estolides produced by Pseudomonas sp 42a2 grown on oleic acid: production and characterization. J. Am. Oil Chem. Soc. 80:859–866
Pym A.S., Brodin P., Brosch R., Huerre M., Cole S.T. (2002) Loss of RD1 contributed to the attenuation of the live tuberculosis vaccines Mycobacterium bovis BCG and Mycobacterium microti. Mol. Microbiol. 46: 709–717
Recht J., Martinez A., Torello S., Kolter R. (2000) Genetic analysis of sliding motility in Mycobacterium smegmatis. J. Bacteriol. 182: 4348–4351
Reddy V.M., Luna-Herrera J., Gangadharam P.R.J. (1996) Pathobiological significance of colony morphology in Mycobacterium avium complex. Microb. Pathog. 21: 97–109
Roach D.R., Martin E., Bean A.G., Rennick D.M., Biscoe H., Britton W.J. (2001) Endogenous inhibition of antimycobacterial immunity by IL-10 varies between mycobacterial species. Scand. J. Immunol. 54: 163–170
Rotzsche H. (1991) Gas chromatographic analysis of fatty acid salts. J. Chromat. A. 552: 281–288
Sacchi R., Addeo F., Paolillo L. (1997) 1H and 13C NMR of virgin olive oil. An overview. Magn. Reson. Chem. 35:S133-S145
Schaefer W.B., Davis C.L., Cohn M.L. (1970) Pathogenicity of transparent, opaque, and rough variants of Mycobacterium avium in chickens and mice. Am. Rev. Respir. Dis. 102: 499–506
Shirtcliffe P.M., Eastophe S.E., Cheng S., Weatherall M., Tan P.L., Le gros G., Beasley R. (2001) The Effect of Delipidated Deglycolipidated (DDMV) and Heat-killed Mycobacterium vaccae in Asthma. Am. J. Respir. Crit. Care Med. 163:1410–1414
Shirtcliffe P.M., Goldkorn A., Weatherall M., Tan P.L., Beasley R. (2003) Pilot study of the safety and effect of intranasal delipidated acid-treated Mycobacterium vaccae in adult asthma. Respirology 8:497–503
Skinner M.A., Prestidge R., Yuan S., Strabala T.J., Tan P.L. (2001) The ability of heat-killed Mycobacterium vaccae to stimulate a cytotoxic T-cell response to an unrelated protein is associated with a 65 kDa heat-shock protein. Immunology 2: 225–233
Skinner M.A., Yuan S., Prestidge R., Chuk D., Watson J.D., Tan P.L.J. (1997) Immunization with heat-killed Mycobacterium vaccae stimulates CD81 cytotoxic T cells specific for macrophages infected with Mycobacterium tuberculosis. Infect. Immun. 65:4525–4530
Stanford J.L., Paul R.C. (1973) A preliminary report on some studies of environmental mycobacteria. Ann. Soc. Belg. Med. Trop. 53: 389–393
Stanford J., Stanford C., Grange J. (2004) Immunotherapy with Mycobacterium vaccae in the treatment of tuberculosis. Front. Biosci. 9: 1701–1719
Thornton A.M., Shevach E.M. (1998) CD4+CD25+ immunoregulatory T cells supress polyclonal T cell activation in vitro by inhibiting interleukin-2 production. J. Exp. Med. 188: 287–296
Van Boxtel R.M., Lambrecht R.S., Collins M.T. (1990) Effects of colonial morphology and tween 80 on antimicrobial susceptibility of Mycobacterium paratuberculosis. Antimicrob. Agents Chemother. 34: 2300–2303
Vestal A.L., Kubica G.P. (1966) Differential colonial characteristics of mycobacteria on Middlebrook and Cohn 7H10 agar–base medium. Am. Rev. Respir. Dis. 94: 247–252
Wang C.C., Rook G.A.W. (1998) Inhibition of an established allergic response to ovalbumin in BALB/c mice by killed Mycobacterium vaccae. Immunology 93: 307–313
Wayne L.G., Kubica G.P. (1986) The mycobacteria. In: Holt J.G., Sneath P.H., Mair N.S., Sharpe M.E. (eds) Bergey’s Manual of Systematic Bacteriology, Vol. 2. Williams & Wilkins, Baltimore, MD, pp 1435–1457
Zuany-Amorim C., Manlius C., Trifilieff C., Brunet L.R., Rook G., Bowen G., Pay G., and Walker C. (2002) Long-term protective and antigen-specific effect of heat-killed Mycobacterium vaccae in a murine model of allergic pulmonary inflammation. J. Immunol. 169:1492–1499
Acknowledgements
This work was supported by the Ministerio de Educación y Cultura (PM98-0180), Ministerio de Ciencia y Tecnología (SAF2002-00514), the Generalitat de Catalunya (2002SGR-00099) and the Fundación Ma Francisca de Roviralta. Esther Julián was recipient of a ‘Beca de Apoyo a la Investigación’ by the Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica.
The authors wish to thank Mercè Martí of the Servei de Microscopia (UAB) for their help in microscopic analysis, and the staff of the Servei d’Immunologia (Hospital Universitari de Bellvitge).
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Rodríguez-Güell, E., Agustí, G., Corominas, M. et al. The production of a new extracellular putative long-chain saturated polyester by smooth variants of Mycobacterium vaccae interferes with Th1-cytokine production. Antonie Van Leeuwenhoek 90, 93–108 (2006). https://doi.org/10.1007/s10482-006-9062-1
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DOI: https://doi.org/10.1007/s10482-006-9062-1