Isolation of recombinant fragments of the major outer-membrane protein of Chlamydia trachomatis: their potential as subunit vaccines

Conlan, J. Wayne; Ferris, Stéphane; Clarke, Ian N.; Ward, Michael E.

doi:10.1099/00221287-136-10-2013

Cited by 14 publications

(4 citation statements)

References 26 publications

(26 reference statements)

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“…The MOMP, which is found in both EBs and RBs, is thought to act as a porin (Bavoil etal., 1984) and has a role in the attachment and uptake of EBs by host cells. It is also recognized as an exposed surface molecule possessing infection-neutralizing epitopes (Collett e t al., 1989 ;Zhang e t al., 1989 ;Conlan et al, 1989 ;Conlan et al, 1990;Peterson e t al., 1990). Polyacrylamide gel analysis of C. tracbomatis proteins has shown that the 60 kDa CrP occurs as a singlet in the trachoma biovar and as a 60/62 kDa doublet in the LGV biovar (Batteiger e t d., 1985).…”

Section: Members Of the Genusmentioning

confidence: 99%

“…Purified chlamydiai preparations from each of the three species that were a mixture of EB with intermediate bodies and a few reticulate bodies were used in these studies. An antiserum raised to C. trachomatis recombinant MOMP expressed in E. coli as insoluble cytoplasmic inclusion bodies (Conlan et al, 1990) was used as a positive control to demonstrate the technique. The results indicated that none of the antisera to the various 60 kDa CrPs showed any surface labelling in either heterologous or homologous combinations.…”

Section: Surface Accessibility Of 60 Kda Crp Epitopesmentioning

confidence: 99%

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Immunoreactivity of the 60 kDa cysteine-rich proteins of Chlamydia trachomatis, Chlamydia psittaci and Chlamydia pneumoniae expressed in Escherichia coli

et al. 1994

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The 60 kDa cysteine-rich proteins (CrPs) of Chlamydia are developmentally regulated outer envelope proteins synthesized late in the chlamydial growth cycle. These proteins, found only on the extracellular infectious elementary bodies, elicit major antibody responses in chlamydial infection. We have cloned and expressed in Escherichia coli the complete 60 kDa CrP genes from Chlamydia trachomatis, C. psittaci and C. pneumoniae. The recombinant products were expressed as either 'native' proteins or as fusions with the bacteriophage T7 gene 10 protein. Electron microscopy showed that recombinant proteins were produced as insoluble inclusions within the E. coli host cells. The recombinant 60 kDa CrPs were purified and used to raise high titre polyclonal antisera. In immunoblot analysis these antisera reacted with the 60 kDa CrPs from purified elementary bodies of all three chlamydial species in a genus-specif ic manner. Further molecular analysis allowed the genus-specif ic cross-reacting epitopes to be localized by using overlapping synthetic peptides covering the C. trachomatis 60 kDa CrP. lmmunogold labelling experiments using purified infectious elementary bodies from the three chlamydial species indicated that the 60 kDa CrPs are not surface accessible to antibody binding.

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Section: Members Of the Genusmentioning

confidence: 99%

Section: Surface Accessibility Of 60 Kda Crp Epitopesmentioning

confidence: 99%

Immunoreactivity of the 60 kDa cysteine-rich proteins of Chlamydia trachomatis, Chlamydia psittaci and Chlamydia pneumoniae expressed in Escherichia coli

et al. 1994

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“…Prior vaccination experiments conducted in animal models of either C. trachomatis or C. psittaci infections have used purified inactivated elementary bodies (EB), purified MOMP or recombinant MOMP (rMOMP) expressed by Escherichia coli, and subfraction or subunit preparations of MOMP (2,5,9,12,17,19). However, these methods had substantial limitations that would be overcome if the outer membrane protein 1 (omp1) gene encoding the MOMP could be expressed in the host cell itself.…”

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confidence: 99%

High-Level Expression ofChlamydia psittaciMajor Outer Membrane Protein in COS Cells and in Skeletal Muscles of Turkeys

et al. 1998

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The omp1 genes encoding the major outer membrane proteins (MOMPs) of avian Chlamydia psittaci serovar A and D strains were cloned and sequenced. The nucleotide sequences of the avian C. psittaci serovar A and D MOMP genes were found to be 98.9 and 87.8% identical, respectively, to that of the avianC. psittaci serovar A strain 6BC, 84.6 and 99.8% identical to that of the avian C. psittaci serovar D strain NJ1, 79.1 and 81.1% identical to that of the C. psittaci guinea pig inclusion conjunctivitis strain, 60.9 and 62.5% identical to that of the Chlamydia trachomatis L2 strain, and 57.5 and 60.4% identical to that of the Chlamydia pneumoniae IOL-207 strain. The serovar A or D MOMPs were cloned in the mammalian expression plasmid pcDNA1. When pcDNA1/MOMP A or pcDNA1/MOMP D was introduced into COS7 cells, a 40-kDa protein that was identical in size, antigenicity, and electrophoretic mobility to native MOMP was produced. Recombinant MOMP (rMOMP) was located in the cytoplasm of transfected COS7 cells as well as in the plasma membrane and was immunoaccessible. Intramuscular administration of pcDNA1/MOMP in specific-pathogen-free turkeys resulted in local expression of rMOMP in its native conformation, after which anti-MOMP antibodies appeared in the serum.

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“…These studies were based on the simple notion that neutralizing antibodies to surface-exposed antigenic sites on the organism constituted immunity. Great efforts were devoted to investigating antibody responses following immunization or infection and on localization and characterization of neutralization epitope(s) on chlamydial antigens (5)(6)(7)(8)(9)(10)(11)(12)(13). The data that supported the notion of antibodymediated immunity arose from observations that serum and monoclonal antibodies specific for C trachomatis are able to neutralize the organism in tissue culture (14,15).…”

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confidence: 99%

T Lymphocyte Immunity in Host Defence against Chlamydia trachomatis and Its Implication for Vaccine Development

Yang

Brunham

1997

Canadian Journal of Infectious Diseases and Medical Microbiolog

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Chlamydia trachomatis is an obligate intracellular bacterial pathogen that causes several significant human infectious diseases, including trachoma, urethritis, cervicitis and salpingitis, and is an important cofactor for transmission of human immunodeficiency virus. Until very recently, over three decades of research effort aimed at developing a C trachomatis vaccine had failed, due mainly to the lack of a precise understanding of the mechanisms for protective immunity. Although most studies concerning protective immunity to C trachomatis have focused on humoral immune responses, recent studies have clearly shown that T helper-1 (Th1)-like CD4 T cell-mediated immune responses play the dominant role in protective immunity. These studies suggest a paradigm for chlamydial immunity and pathology based on the concept of heterogeneity (Th1/Th2) in CD4 T cell immune responses. This concept for chlamydial immunity offers a rational template on which to base renewed efforts for development of a chlamydial vaccine that targets the induction of cell-mediated Th1 immune responses.

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Isolation of recombinant fragments of the major outer-membrane protein of Chlamydia trachomatis: their potential as subunit vaccines

Cited by 14 publications

References 26 publications

Immunoreactivity of the 60 kDa cysteine-rich proteins of Chlamydia trachomatis, Chlamydia psittaci and Chlamydia pneumoniae expressed in Escherichia coli

Immunoreactivity of the 60 kDa cysteine-rich proteins of Chlamydia trachomatis, Chlamydia psittaci and Chlamydia pneumoniae expressed in Escherichia coli

High-Level Expression ofChlamydia psittaciMajor Outer Membrane Protein in COS Cells and in Skeletal Muscles of Turkeys

T Lymphocyte Immunity in Host Defence against Chlamydia trachomatis and Its Implication for Vaccine Development

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