Immunotherapy with fragmented Mycobacterium tuberculosis cells increases the effectiveness of chemotherapy against a chronical infection in a murine model of tuberculosis

doi:10.1016/j.vaccine.2004.09.008

Vaccine

Volume 23, Issue 11, 3 February 2005, Pages 1393-1398

https://doi.org/10.1016/j.vaccine.2004.09.008 Get rights and content

Abstract

Reduction of colony forming units by rifampicin-isoniazid therapy given 9–17 weeks post-infection was made more pronounced by immunotherapy with a vaccine made of fragmented Mycobacterium tuberculosis cells detoxified and liposomed (RUTI), given on weeks 17, 19 and 21 post-infection, in the murine model of tuberculosis in C57BL/6 and DBA/2 inbred strains. RUTI triggered a Th1/Th2 response, as demonstrated by the production of IgG1, IgG2a and IgG3 antibodies against a wide range of peptides. The histological analysis did not show neither eosinophilia nor necrosis, and granulomatous infiltration was only slightly increased in C57BL/6 mice when RUTI was administered intranasally.

Introduction

It has been estimated that one third of mankind harbours a latent infection with Mycobacterium tuberculosis [1]. Latent bacilli have to adapt to stressful conditions generated by the infected host against them by slowing down the metabolism or becoming dormant [2], [3]. Since chemotherapy and immunity are mainly directed against growing bacilli [4], [5], their destruction in a short period of time is difficult. It has been hypothesized the usefulness of a immunotherapeutic vaccine, but the results of some recent studies strongly recommend not to use post-infection vaccination with killed or alive BCG, as this induces strong tissue toxicity [6], [7]. In the present study, we have assessed the benefit of the administration of a fragmented M. tuberculosis cells formulation (RUTI) to reduce the bacterial load after the treatment with chemotherapy in a murine model of chronical tuberculosis infection.

Section snippets

Experimental infection

Specific pathogen-free DBA/2 and C57BL/6 female mice, 6 weeks old, were provided by Charles River (L’Arbresle Cedex, France) and kept under controlled conditions in a P3 High Security Facility with sterile food and water ad libitum. M. tuberculosis standard strain NC007416 (H37Rv) was grown in Proskauer Beck medium containing 0.01% Tween 80 to mid-log phase and stored in aliquots at −70 °C. Mice were placed in a Middlebrook Airbone Infection Apparatus (Glas-col Inc., Terre Haute, IN, USA). The

Results

Fig. 2 shows the bactericidal activity induced by the inoculation of RUTI IN or SC in both susceptible or resistant mice strains [8], [11] after chemotherapy, which also impedes the re-activation suffered by mice treated only with chemotherapy. Besides, inoculation of RUTI without a previous chemotherapy did not affect the bacterial load although elicited a similar immunological response (data not shown). This bactericidal activity seems to be related to the Th1 response as it is demonstrated

Discussion

RUTI might be a useful vaccine for reducing the period of chemotherapy in M. tuberculosis infection at the point when it has less efficacy because of the presence of latent bacilli in the lesions [13], [14], [15], [16]. Its mechanism of action might be related to the induction of a Th1 response against a wide range of M. tuberculosis antigen able to elicit a protective response against M. tuberculosis infection like ESAT-6, CFP-10, Ag85 or Hsp70 [4]. Also, its effectiveness is necessarily

Acknowledgement

This study was supported by Grant FIS 01/3104 of the Spanish Ministry of Health. Data were partially presented as a communication in the Experimental Biology Congress in Washington DC (USA) on April 2004.

References (28)

J. Grosset
Bacteriologic basis of short-course chemotherapy for tuberculosis
Clin Chest Med
(1980)
S. Cobbold et al.
Infectious tolerance
Curr Opin Immunol
(1998)
V. Gruppo et al.
Dose of BCG does not influence the efficient generation of protective immunity in mice challenged with Mycobacterium tuberculosis
Tuberculosis (Edinb)
(2002)
T.E. Karplus et al.
A new method for reduction of endotoxin contamination from protein solutions
J Immunol Methods
(1987)
World Health Organization. Global Tuberculosis Control. WHO Report 2001, Geneva, Switzerland,...
A.F. Cunningham et al.
Mycobacterial stationary phase induced by low oxygen tension: cell wall thickening and localization of the 16-kilodalton alpha-crystallin homolog
J Bacteriol
(1998)
T.M. Michele et al.
Exposure to antibiotics induces expression of the Mycobacterium tuberculosis sigF gene: implications for chemotherapy against mycobacterial persistors
Antimicrob Agents Chemother
(1999)
P. Andersen et al.
T-cell proliferative response to antigens secreted by Mycobacterium tuberculosis
Infect Immun
(1991)
A.L. Moreira et al.
Mycobacterial antigens exacerbate disease manifestations in Mycobacterium tuberculosis-infected mice
Infect Immun
(2002)
J. Turner et al.
Effective preexposure tuberculosis vaccines fail to protect when they are given in an immunotherapeutic mode
Infect Immun
(2000)

P.J. Cardona et al.

Widespread bronchogenic dissemination makes DBA/2 mice more susceptible than C57BL/6 mice to experimental aerosol infection with Mycobacterium tuberculosis

Infect Immun

(2003)

M.G. Sonnenberg et al.

Definition of Mycobacterium tuberculosis culture filtrate proteins by two-dimensional polyacrylamide gel electrophoresis, N-terminal amino acid sequencing, and electrospray mass spectrometry

Infect Immun

(1997)

T. Rabilloud

Mechanisms of protein silver staining in polyacrylamide gels: a 10-year synthesis

Electrophoresis

(1990)

E. Medina et al.

Resistance ranking of some common inbred mouse strains to Mycobacterium tuberculosis and relationship to major histocompatibility complex haplotype and Nramp1 genotype

Immunology

(1998)

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Immunotherapy with fragmented Mycobacterium tuberculosis cells increases the effectiveness of chemotherapy against a chronical infection in a murine model of tuberculosis

Abstract

Introduction

Section snippets

Experimental infection

Results

Discussion

Acknowledgement

Clin Chest Med

Curr Opin Immunol

Tuberculosis (Edinb)

J Immunol Methods

Mycobacterial stationary phase induced by low oxygen tension: cell wall thickening and localization of the 16-kilodalton alpha-crystallin homolog

J Bacteriol

Exposure to antibiotics induces expression of the Mycobacterium tuberculosis sigF gene: implications for chemotherapy against mycobacterial persistors

Antimicrob Agents Chemother

T-cell proliferative response to antigens secreted by Mycobacterium tuberculosis

Infect Immun

Mycobacterial antigens exacerbate disease manifestations in Mycobacterium tuberculosis-infected mice

Infect Immun

Effective preexposure tuberculosis vaccines fail to protect when they are given in an immunotherapeutic mode

Infect Immun

Widespread bronchogenic dissemination makes DBA/2 mice more susceptible than C57BL/6 mice to experimental aerosol infection with Mycobacterium tuberculosis

Infect Immun

Definition of Mycobacterium tuberculosis culture filtrate proteins by two-dimensional polyacrylamide gel electrophoresis, N-terminal amino acid sequencing, and electrospray mass spectrometry

Infect Immun

Mechanisms of protein silver staining in polyacrylamide gels: a 10-year synthesis

Electrophoresis

Resistance ranking of some common inbred mouse strains to Mycobacterium tuberculosis and relationship to major histocompatibility complex haplotype and Nramp1 genotype

Immunology