scholarly journals Immunogenicity Studies with Microbial Fractions of M. tuberculosis H37Rv Total Culture Filtrate

2015 ◽  
Vol 8 (1) ◽  
pp. 48
Author(s):  
Rajnish Kumar ◽  
Parminder Jit Kaur ◽  
G. K. Khuller ◽  
Indu Verma

<p class="1Body">Current study investigates the whole secretory proteome of <em>Mycobacterium tuberculosis</em> as culture filtrate fractions to identify immunoprotective protein antigens on the basis of protection studies in animal (mouse and guinea pig) models. Secretory culture filtrate proteins (CFPs) of <em>M. tuberculosis</em> H<sub>37</sub>Rv were fractionated into fifteen narrow molecular mass fractions in the order of increasing molecular size (F1-F15) by electroelution. Immunization studies revealed proteins in the molecular weight range of 20-24kDa (F7), 25-30kDa (F8) and 37-42kDa (F11) as key protective fractions against experimental tuberculosis in both the animal (mice and guinea pig) models. Amongst these fractions, F7 imparted even better protection as compared to BCG. Immunological studies with all the fractions demonstrated that although selected three protective fractions were able to induce significant immune responses in both short term culture filtrate (STCF) immunized and Mtb infected animals, there were number of other non-protective fractions also that were inducing higher immune responses either in immunized animals (e.g.F12-F15) or in Mtb challenged animals (e.g.F1-F6). These results demonstrate that only those mycobacterial proteins that are recognized by the host immune system both during immunization and infection can induce significant protection against experimental tuberculosis, however there is no direct correlation between the level of immune responses and degree of protective efficacy.</p>

2000 ◽  
Vol 68 (10) ◽  
pp. 5809-5815 ◽  
Author(s):  
D. Neil Wedlock ◽  
Bridget Vesosky ◽  
Margot A. Skinner ◽  
Geoffrey W. de Lisle ◽  
Ian M. Orme ◽  
...  

ABSTRACT In this study vaccines prepared from culture filtrate proteins (CFP) of Mycobacterium bovis and interleukin-2 (IL-2) were tested in cattle for their capacity to stimulate immune responses and to protect against an intratracheal challenge with virulent M. bovis. Nine groups of cattle were vaccinated with combinations of different doses of CFP and bovine IL-2 mixed with a monophosphoryl lipid A (MPL) adjuvant. An additional group was vaccinated withM. bovis BCG. Immune responses in CFP–IL-2-vaccinated animals differed from those seen in BCG-vaccinated animals by inducing high antigen-specific antibody responses and low levels of gamma interferon and IL-2 released from purified protein derivative-stimulated whole-blood cultures. In a concurrent experiment, additional animals were added to the high-dose CFP–IL-2, MPL control, and BCG groups and these expanded groups of animals were challenged intratracheally with virulent M. bovis. Although the lung lesion scores were significantly lower for both the CFP–IL-2-and BCG-vaccinated groups compared to the MPL control group, the overall level of protection was greatest for the BCG-vaccinated animals. There were more animals with extrathoracic spread of disease in the CFP–IL-2 group than in the other groups. While vaccination of cattle withM. bovis CFP gave an encouraging reduction in tuberculous lesions and did not induce a delayed-type hypersensitivity response to PPD, future CFP vaccines must prevent any extrathoracic spread of disease.


Vaccine ◽  
2012 ◽  
Vol 30 (43) ◽  
pp. 6198-6209 ◽  
Author(s):  
Ankan Gupta ◽  
F.J. Ahmad ◽  
Faiz Ahmad ◽  
U.D. Gupta ◽  
M. Natarajan ◽  
...  

mBio ◽  
2020 ◽  
Vol 11 (2) ◽  
Author(s):  
Tania Rivera-Hernandez ◽  
Mira Syahira Rhyme ◽  
Amanda J. Cork ◽  
Scott Jones ◽  
Celia Segui-Perez ◽  
...  

ABSTRACT Recent global advocacy efforts have highlighted the importance of development of a vaccine against group A Streptococcus (GAS). Combo5 is a non-M protein-based vaccine that provides protection against GAS skin infection in mice and reduces the severity of pharyngitis in nonhuman primates. However, Combo5 with the addition of aluminum hydroxide (alum) as an adjuvant failed to protect against invasive GAS infection of mice. Here, we show that formulation of Combo5 with adjuvants containing saponin QS21 significantly improves protective efficacy, even though all 7 adjuvants tested generated high antigen-specific IgG antibody titers, including alum. Detailed characterization of Combo5 formulated with SMQ adjuvant, a squalene-in-water emulsion containing a TLR4 agonist and QS21, showed significant differences from the results obtained with alum in IgG subclasses generated following immunization, with an absence of GAS opsonizing antibodies. SMQ, but not alum, generated strong interleukin-6 (IL-6), gamma interferon (IFN-γ), and tumor necrosis alpha (TNF-α) responses. This work highlights the importance of adjuvant selection for non-M protein-based GAS vaccines to optimize immune responses and protective efficacy. IMPORTANCE Availability of a group A Streptococcus vaccine remains an unmet public health need. Here, we tested different adjuvant formulations to improve the protective efficacy of non-M protein vaccine Combo5 in an invasive disease model. We show that novel adjuvants can dramatically shape the type of immune response developed following immunization with Combo5 and significantly improve protection. In addition, protection afforded by Combo5 is not mediated by opsonizing antibodies, believed to be the main correlate of protection against GAS infections. Overall, this report highlights the importance of adjuvant selection in raising protective immune responses against GAS invasive infection. Adjuvants that can provide a more balanced Th1/Th2-type response may be required to optimize protection of GAS vaccines, particularly those based on non-M protein antigens.


1994 ◽  
Vol 62 (8) ◽  
pp. 3454-3462 ◽  
Author(s):  
R Weeratna ◽  
D A Stamler ◽  
P H Edelstein ◽  
M Ripley ◽  
T Marrie ◽  
...  

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Mandi Liu ◽  
Yue Zhang ◽  
Di Zhang ◽  
Yun Bai ◽  
Guomei Liu ◽  
...  

AbstractEnterotoxigenic Escherichia coli (ETEC), an essential cause of post-weaning diarrhea (PWD) in piglets, leads to significant economic losses to the pig industry. The present study aims to identify the role of ETEC total RNA in eliciting immune responses to protect animals against ETEC infection. The results showed that the total RNA isolated from pig-derived ETEC K88ac strain effectively stimulated the IL-1β secretion of porcine intestinal epithelial cells (IPEC-J2). The mouse model immunized with ETEC total RNA via intramuscular injection (IM) or oral route (OR) was used to evaluate the protective efficiency of the ETEC total RNA. The results suggested that 70 μg ETEC total RNA administered by either route significantly promoted the production of the serum IL-1β and K88ac specific immunoglobulins (IgG, IgM, and IgA). Besides, the ETEC RNA administration augmented strong mucosal immunity by elevating K88ac specific IgA level in the intestinal fluid. Intramuscularly administered RNA induced a Th1/Th2 shift toward a Th2 response, while the orally administered RNA did not. The ETEC total RNA efficiently protected the animals against the ETEC challenge either by itself or as an adjuvant. The histology characterization of the small intestines also suggested the ETEC RNA administration protected the small intestinal structure against the ETEC infection. Particularly of note was that the immunity level and protective efficacy caused by ETEC RNA were dose-dependent. These findings will help understand the role of bacterial RNA in eliciting immune responses, and benefit the development of RNA-based vaccines or adjuvants.


1996 ◽  
Vol 24 (1) ◽  
pp. 73-79
Author(s):  
Laura S. Babcock ◽  
Thomas T. Kawabata ◽  
Victor S. Moore ◽  
Catherine M. Condo ◽  
Annette Prentø

Specific guinea-pig IgG1 has traditionally been measured by using an in vivo guinea-pig passive cutaneous anaphylaxis (PCA) assay. This paper describes the development and validation of a quantitative enzyme-linked immunosorbent assay (ELISA) for specific IgG1 against two protein antigens (Alcalase and Enzyme B) as an alternative to the PCA assay. The ELISA format involved a rabbit antibody bound to microtitre plates to capture the antigen. The test sera is added to this, followed sequentially by goat anti-guinea-pig IgG1 and rabbit anti-goat-IgG-alkaline phosphatase conjugate. Aliquots of each serum sample from immunised guinea-pigs were analysed with the ELISA for specific IgG1 titres in three laboratories and were compared to titres determined by using the PCA assay. The findings demonstrate that there is a good correlation between the ELISA and the PCA assay and that the ELISA shows good interlaboratory reproducibility. Thus, the antibody capture ELISA described in this report is a valid and robust replacement for the guinea-pig PCA assay.


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