Naive Human T Cells Develop into Th1 Effectors after Stimulation with<i>Mycobacterium tuberculosis</i>-Infected Macrophages or Recombinant Ag85 Proteins

Russo, Donna; Kozlova, N. M.; Lakey, David; Kernodle, Douglas S.

doi:10.1128/iai.68.12.6826-6832.2000

Cited by 20 publications

(17 citation statements)

References 39 publications

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“…Despite past studies demonstrating that T cells are pivotal in the progression of mycoplasma respiratory disease (11), little is known about the T cell responses and their association with pathogenesis of any mycoplasma disease. In particular, the activation of Th1 and Th2 cell subsets has not been examined in any mycoplasma disease, despite their impact in the progression and pathology of other diseases (27)(28)(29)(30)(31)(32)(33)(34)(35). Because of the critical role that T cell responses have in modulating immunity, it is important to ascertain which T cell populations are responding to the mycoplasma infection and their contribution to disease pathogenesis.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, Th2 cells control Ab responses through their secretion of characteristic cytokines (e.g., IL-4). Studies have shown that Th cell subset activation is important in the control of other infectious diseases (27)(28)(29)(30)(31)(32)(33)(34)(35), but their impact on mycoplasma respiratory disease is unknown. In addition, CD8 ϩ T cells are also known to impact on the progression of inflammatory diseases, as they can regulate these responses (36,37) or cause lung injury through CD8 ϩ T cell-mediated apoptosis (38 -40).…”

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confidence: 99%

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Depletion of CD8+ T Cells Exacerbates CD4+ Th Cell-Associated Inflammatory Lesions During Murine Mycoplasma Respiratory Disease

Jones

Tabor

Sun

et al. 2002

The Journal of Immunology

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Mycoplasma infection is a leading cause of pneumonia worldwide and can lead to other respiratory complications. A component of mycoplasma respiratory diseases is immunopathologic, suggesting that lymphocyte activation is a key event in the progression of these chronic inflammatory diseases. The present study delineates the changes in T cell populations and their activation after mycoplasma infection and determines their association with the pathogenesis of murine Mycoplasma respiratory disease, due to Mycoplasma pulmonis infection. Increases in T cell population numbers in lungs and lower respiratory lymph nodes were associated with the development of mycoplasma respiratory disease. Although both pulmonary Th and CD8+ T cells increased after mycoplasma infection, there was a preferential expansion of Th cells. Mycoplasma-specific Th2 responses were dominant in lower respiratory lymph nodes, while Th1 responses predominated in spleen. However, both mycoplasma-specific Th1 and Th2 cytokine (IL-4 and IFN-γ) responses were present in the lungs, with Th1 cell activation as a major component of the pulmonary Th cell response. Although a smaller component of the T cell response, mycoplasma-specific CD8+ T cells were also a significant component of pulmonary lymphoid responses. In vivo depletion of CD8+ T cells resulted in dramatically more severe pulmonary disease, while depletion of CD4+ T cells reduced its severity, but there was no change in mycoplasma numbers in lungs after cell depletion. Thus, mycoplasma-specific Th1 and CD8+ T cell activation in the lung plays a critical regulatory role in development of immunopathologic reactions in Mycoplasma respiratory disease.

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

Depletion of CD8+ T Cells Exacerbates CD4+ Th Cell-Associated Inflammatory Lesions During Murine Mycoplasma Respiratory Disease

Jones

Tabor

Sun

et al. 2002

The Journal of Immunology

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“…There are several reports describing the cloning and expression of the CFP-10 gene in different expression vectors such as pUC18 (12), pRSETB (13), pGEMt-Easy vector, pVAX1 (14), pMCT6 (15) and pET17b (16). In a series of studies Lakey et al, reported cloning and expression of Ag85A and Ag85B.…”

Section: Discussionmentioning

confidence: 99%

“…To clone this gene, they digested plasmid vector and performed insertion with restriction enzyme KpnΙ (12). Similarly, other researchers used this technique (double cut) to clone this gene (13,15,16).…”

Section: Discussionmentioning

confidence: 99%

Cloning and Expression of Mycobacterium tuberculosis Major Secreted Protein Antigen 85B (Ag85B) in Escherichia coli

Zarif

Sankian

Gholubi

et al. 2013

Jundishapur J Microbiol

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Background:The 30 kDa major secretory protein of Mycobacterium tuberculosis (antigen 85B) is a primary vaccine candidate. This secreted antigen induces a protective immune response and stimulates the production of IFN-γ in animal models. Objectives: The aim of this study was cloning and expression of Ag 85B of M. tuberculosis in Escherichia coli. Materials and Methods: To produce recombinant Ag85B, the fbpB gene was amplified by PCR method. Then inserted into the pET101/D vector and transported into E. coli strain TOPO10. Plasmid containing pET101/D: Ag85B was transformed into competence E.coli BL21 (DE3). The transformed E.coli strain BL21 was effectively expressed recombinant Ag85B. Results: The expressed fusion protein was found almost entirely in the insoluble form. Followed by sonication to disrupt the cells, Solution of the cell debris was centrifuged and after use of Ni-NTA column and 6 molar urea and 6 M guanidine-HCl solutions recombinant protein was purified. Conclusions: These results could serve as a basis for further studies in endemic regions of tuberculosis on the usefulness of this gene and its expression product in the development of subunit vaccine and DNA vaccine against tuberculosis.

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“…[17][18][19][20][21] The present studies are concerned with the delivery of proteins for local activity, specifically the delivery of subunit vaccines to elicit cell-mediated responses for tuberculosis prevention. 22 Many of these proteins are in limited supply and a formulation strategy requires an approach validated using model proteins. The enzyme, β-glucuronidase (GUS), was evaluated as a model protein in preliminary studies of liposome formulation, lyophilization, micronization, and aerodynamic size distributions.…”

Section: Introductionmentioning

confidence: 99%

Liposomal dry powders as aerosols for pulmonary delivery of proteins

Hickey

2005

AAPS PharmSciTech

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The purpose of this research was to develop liposomal dry powder aerosols for protein delivery. The delivery of stable protein formulations is essential for protein subunit vaccine delivery, which requires local delivery to macrophages in the lungs. β-Glucuronidase (GUS) was used as a model protein to evaluate dry powder liposomes as inhaled delivery vehicles. Dimyristoyl phosphatylcholine:cholesterol (7:3) was selected as the liposome composition. The lyophilization of liposomes, micronization of the powders, aerosolization using a dry powder inhaler (DPI), and in vitro aerodynamic fine particle fraction upon collection in a twinstage liquid impinger were evaluated. After lyophilization and jet-milling, the total amount of GUS and its activity, representing encapsulation efficiency and stability, were evaluated. The GUS amount and activity were measured and compared with freshly-prepared liposomes in the presence of mannitol, 43% of initial GUS amount, 29% of GUS activity after lyophilization and 36% of GUS amount, 22% of activity after micronization were obtained. Emitted doses from dry powder inhaler were 53%, 58%, 66%, and 73% for liposome powder:mannitol carrier ratios of 1:0, 1:4, 1:9, and 1:19. Fifteen percent of the liposome particles were less than 6.4 μm in aerodynamic diameter. The results demonstrate that milled liposome powders containing protein molecules can be aerosolized effectively at a fixed flow rate. Influences of different cryoprotectants on lyophilization of protein liposome formulations are reported. The feasibility of using liposomal dry powder aerosols for protein delivery has been demonstrated but further optimization is required in the context of specific therapeutic proteins.

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Naive Human T Cells Develop into Th1 Effectors after Stimulation withMycobacterium tuberculosis-Infected Macrophages or Recombinant Ag85 Proteins

Abstract: Most studies of human T-cell responses in tuberculosis have

Cited by 20 publications

References 39 publications

Depletion of CD8+ T Cells Exacerbates CD4+ Th Cell-Associated Inflammatory Lesions During Murine Mycoplasma Respiratory Disease

Depletion of CD8+ T Cells Exacerbates CD4+ Th Cell-Associated Inflammatory Lesions During Murine Mycoplasma Respiratory Disease

Cloning and Expression of Mycobacterium tuberculosis Major Secreted Protein Antigen 85B (Ag85B) in Escherichia coli

Liposomal dry powders as aerosols for pulmonary delivery of proteins

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