scholarly journals PROTEIN REKOMBINAN 38 KDA MYCOBAKTERIUM TUBERKULOSIS DAPAT MENGIMBAS PEMBUATAN INTERLEUKIN-2 DAN INTERFERON-γ LIMFOSIT T DI KULTUR SEL MONONUKLEAR DARAH TEPI

2018 ◽  
Vol 22 (2) ◽  
pp. 151
Author(s):  
Maimun Z Arthamin ◽  
Singgih Pujo Wahono ◽  
Antiek Primardianti ◽  
Ati Rastini ◽  
Tri Wahju Astuti ◽  
...  
Keyword(s):  
Immune Response ◽  
T Lymphocytes ◽  
Recombinant Protein ◽  
Vaccine Development ◽  
Interleukin 2 ◽  
Development Program ◽  
In Vitro Study ◽  
Cell Mediated Immunity ◽  
Ifn Γ

Tuberculosis (TB) is caused by Mycobacterium tuberculosis (M.tb) and is one of the significant mortality causes WHO (2012). Theprimary immune response in TB pathogenesis is Cell Mediated Immunity (CMI), roled by T lymphocytes. Interleukin-2 (IL-2) is a growthfactor for T lymphocytes. Gamma Interferon is the key cytokine in M.tb infection control, synthezised by T lymphocytes. An effectivevaccination strategy is achieved by giving vaccine which is able to stimulate T lymphocytes in synthezising cytokines. The 38 kDa M.tbprotein is potential in the vaccine development program, because it has specific epitopes for T lymphocytes. The aim of this study was toknow how to determine that the 38 kDa recombinant protein of M.tb Malang strain could induce cellular immune response by IL-2 andIFN-γ synthezised by T lymphocytes. The study was carried out by an experimental in vitro study on PBMC from healthy endemic subjects,those having TB contact, and the TB patients themselves. PBMC from subjects was cultured with 38 kDa recombinant protein of M.tbMalang strain, with PPD and without any protein. The analysis of IL-2 and IFN-γ used flowcytometry. The result showed that the highestpercentage of IL-2 was found in the culture with 38 kDa recombinant protein of M.tb Malang strain, in healthy endemic (p=0.000)and in those who had TB contact (p=0.000). the highest percentage of IFN-γ was found in the culture with 38 kDa recombinant proteinof M.tb Malang strain, in healthy endemic (p=0.007) and those who had TB contact (p = 0.105). The 38 kDa recombinant proteinof M.tb Malang strain was able to induce IL-2 and IFN-γ synthezised by TCD3+ lymphocytes from healthy endemic subjects and thosewho had TB contact.

scholarly journals Gingyo-San Enhances Immunity and Potentiates Infectious Bursal Disease Vaccination

2011 ◽  
Vol 2011 ◽  
pp. 1-10 ◽  
Author(s):  
Che-Ming Hung ◽  
Chia-Chou Yeh ◽  
Kowit-Yu Chong ◽  
Hsiao-Ling Chen ◽  
Jiun-Yu Chen ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Interleukin 2 ◽  
Serum Antibody ◽  
Infectious Bursal Disease ◽  
Interleukin 4 ◽  
Interleukin 12 ◽  
Cell Mediated Immunity ◽  
Bursal Disease ◽  
Antibody Titers ◽  
Ifn Γ

The purpose of the present study was to investigate the effects of Gingyo-san (GGS), a traditional Chinese medical formula, on peripheral lymphocyte proliferation and serum antibody titers in chickens vaccinated against the infectious bursal disease (IBD) virus. Treatment groups were fed one of three doses of GGS in their diet (0.5%, 1.0% and 2.0%, w/w), and the IBD vaccine was administered at 1 and 3 weeks of age. At Weeks 8, 12 and 16, changes in serum IBD antibody titers were measured via the micro-method and T cell proliferation. In gene expression experiments, GGS-treated peripheral T lymphocytes were stimulated with concanavalin A (ConA) for 24 h. The mRNA expression of interleukin-2 (IL-2), interferon-γ (IFN-γ), interleukin-4 (IL-4) and interleukin-12 (IL-12) was determined using a semi-quantitative RT-PCR assay. The results showed that a low dose of GGS could significantly raise the antibody titers. Medium and high doses of GGS enhanced IL-2 and IFN-γ production. GGS altered the expression of IL-4 and IL-12 in T lymphocytes. CD4+T lymphocyte development was also skewed towards the Th1 phenotype. GGS enhanced cell-mediated immunity and augmented the effects of IBD vaccination in strengthening subsequent anti-viral responses.

scholarly journals In Vitro Cell-Mediated Immune Responses of Human Immunodeficiency Virus-Infected and -Uninfected Individuals to Whole Cytomegalovirus Antigens and Their Subunits

2008 ◽  
Vol 15 (9) ◽  
pp. 1398-1409 ◽  
Author(s):  
A. Weinberg ◽  
J. Spritzler ◽  
M. Nokta ◽  
R. Schrier ◽  
A. Landay ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Interleukin 2 ◽  
Cell Mediated Immunity ◽  
Hiv Replication ◽  
Hiv Rna ◽  
Highly Sensitive ◽  
Immunodeficiency Virus ◽  
Ifn Γ ◽  
Cd4 Cell

ABSTRACT The aim of this study was to optimize the ability to detect cytomegalovirus (CMV)-specfic cell-mediated immunity (CMI) in human immunodeficiency virus (HIV)-infected individuals by comparing different assays (the lymphocyte proliferation assay [LPA] and assays for gamma interferon [IFN-γ] and interleukin-2 [IL-2] production) and CMV antigenic preparations. Thresholds discriminating positive from negative CMI results were developed with specimens from 36 CMV-seropositive and 21 CMV-seronegative healthy individuals. The analysis showed that the CMI elicited by any of the four CMV whole lysates tested in this study tended to be more robust and sensitive than the responses to the subunit antigens gB and pp65. LPA and inducible IFN-γ but not IL-2 were highly sensitive measures of CMV-specific CMI in HIV-infected and -uninfected individuals. The ability to detect CMV-specific LPA or IFN-γ responses in HIV-infected individuals significantly increased with higher CD4 cell numbers. Nevertheless, the proportion of HIV-infected subjects with CD4 counts of ≥500 cells/μl who had a detectable CMV-specific CMI remained significantly lower than that of healthy adults. The ability to detect CMV-specific CMI in HIV-infected individuals decreased with higher levels of HIV replication, with discriminative thresholds of 103 to 104 HIV RNA copies/ml of plasma, for LPA or inducible IFN-γ production elicited by different antigens. The LPA responses obtained with CMV whole lysate and phytohemagglutinin were significantly correlated in HIV-infected subjects but not uninfected controls, indicating a novel characteristic of the CMI defect caused by HIV. The intrasubject variabilities of the CMV-specific CMI were similar in HIV-infected and -uninfected individuals. These data show that LPA and the inducible IFN-γ production elicited by CMV whole lysates may be used to assess modifications of the immune competency of HIV-infected individuals.

scholarly journals Induction of Cell-Mediated Immunity to Staphylococcus aureus in the Mouse Mammary Gland by Local Immunization with a Live Attenuated Mutant

2002 ◽  
Vol 70 (8) ◽  
pp. 4254-4260 ◽  
Author(s):  
Marisa I. Gómez ◽  
Daniel O. Sordelli ◽  
Fernanda R. Buzzola ◽  
Verónica E. García
Keyword(s):  
Staphylococcus Aureus ◽  
T Cells ◽  
Mammary Gland ◽  
Mononuclear Cells ◽  
Regional Lymph Node ◽  
Interleukin 2 ◽  
Mouse Mammary Gland ◽  
Cell Mediated Immunity ◽  
Ifn Γ

ABSTRACT The efficacy of intramammary (Ima) immunization with a live attenuated (la) Staphylococcus aureus mutant to protect the mouse mammary gland from infection has previously been established. The present study was aimed at evaluating whether Ima immunization with la-S. aureus can induce cell-mediated immune responses to the pathogen within the mammary gland. Mice were immunized by Ima route with la-S. aureus, and regional lymph node mononuclear cells were obtained thereafter. A higher expression of the interleukin-2 receptor was found on B and T cells from immunized mice when they were compared with control mice. Immunization with la-S. aureus induced strong proliferative responses to S. aureus. Moreover, significantly increased levels of gamma interferon (IFN-γ) were produced by CD4+ T cells when lymphocytes from immunized mice, but not from control mice, were cultured in the presence of staphylococcal antigens. Moreover, a significant increase in the percentage of IFN-γ-producing CD4+ and CD8+ T cells was observed after S. aureus Ima challenge in immunized mice compared to challenged control mice. Our results demonstrated that Ima immunization with la-S. aureus induced primed lymphocyte populations capable of responding against staphylococcal antigens during in vitro stimulation, as well as during in vivo infection by S. aureus. CD4+ and CD8+ T cells appear to be the main lymphocyte subpopulations involved in this response. It is suggested that IFN-γ production induced by Ima immunization may play a pivotal role in the eradication of intracellular staphylococci.

scholarly journals Polyfunctional T Lymphocytes Are in the Peripheral Blood of Donors Naturally Immune to Coccidioidomycosis and Are Not Induced by Dendritic Cells

2009 ◽  
Vol 78 (1) ◽  
pp. 309-315 ◽  
Author(s):  
Lance Nesbit ◽  
Suzanne M. Johnson ◽  
Demosthenes Pappagianis ◽  
Neil M. Ampel
Keyword(s):  
Dendritic Cells ◽  
T Lymphocytes ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
High Expression ◽  
Peripheral Blood Mononuclear ◽  
Necrosis Factor Alpha ◽  
Ifn Γ

ABSTRACT Coccidioidomycosis is a fungal infection endemic in the southwestern United States that is increasing in incidence. While cellular immunity correlates with protection from clinical illness, the precise elements of that response are undefined. Using the coccidioidal antigen preparation T27K and multiparametric flow cytometry, the in vitro frequency of polyfunctional T lymphocytes in the peripheral blood of naturally immune healthy donors and those who were nonimmune was determined. Polyfunctional CD4 lymphocytes, defined as producing intracellular interleukin 2 (IL-2), gamma interferon (IFN-γ), and tumor necrosis factor alpha simultaneously, had a frequency of 137 per 400,000 events among peripheral blood mononuclear cells (PBMC) of immune donors compared to 11 per 400,000 PBMC from nonimmune donors (P = 0.03). When monocyte-derived mature dendritic cells pulsed with T27K (mDCT27K) were used for antigen presentation, the frequency of polyfunctional CD4 T lymphocytes did not significantly increase for either group, although mDCT27K did significantly increase the concentrations of IL-2 and IFN-γ released by PBMC from nonimmune donors (P = 0.02). After in vitro stimulation with T27K, polyfunctional CD4 and CD8 lymphocytes of PBMC from immune donors had a mixture of low- and high-expression CCR7 cells, suggesting both effector and central memory, compared with predominantly high-expression CCR7 cells when PBMC were incubated with the mitogen phytohemagglutinin (P = 0.03). These data demonstrate the presence of polyfunctional T lymphocytes in the peripheral blood of individuals with coccidioidal immunity and suggest a model for the in vitro testing of vaccine candidates for coccidioidomycosis.

scholarly journals Vaccination with Novel Immunostimulatory Adjuvants against Blood-Stage Malaria in Mice

2003 ◽  
Vol 71 (9) ◽  
pp. 5178-5187 ◽  
Author(s):  
Zhong Su ◽  
Mi-Fong Tam ◽  
Dragana Jankovic ◽  
Mary M. Stevenson
Keyword(s):  
Immune Response ◽  
Protective Immunity ◽  
Vaccine Development ◽  
Challenge Infection ◽  
Blood Stage ◽  
Interleukin 12 ◽  
Antigen Preparation ◽  
Malaria Antigen ◽  
Ifn Γ

ABSTRACT An important aspect of malaria vaccine development is the identification of an appropriate adjuvant which is both capable of stimulating a protective immune response and safe for use by humans. Here, we investigated the feasibility of using novel immunostimulatory molecules as adjuvants combined with a crude antigen preparation and coadsorbed to aluminum hydroxide (alum) as a vaccine against blood-stage Plasmodium chabaudi AS malaria. Prior to challenge infection, immunization of genetically susceptible A/J mice with the combination of malaria antigen plus recombinant interleukin-12 (IL-12) in alum induced a Th1 immune response with production of high levels of gamma interferon (IFN-γ) and diminished IL-4 levels by spleen cells stimulated in vitro with parasite antigen compared to mice immunized with antigen alone, antigen in alum, or antigen plus IL-12. Mice immunized with malaria antigen plus recombinant IL-12 in alum had high levels of total malaria-specific antibody and immunoglobulin G2a. Compared to unimmunized mice, immunization with antigen plus IL-12 in alum induced the highest level of protective immunity against challenge infection with P. chabaudi AS, which was evident as a significantly decreased peak parasitemia level and 100% survival. Protective immunity was dependent on CD4+ T cells, IFN-γ, and B cells and was long-lasting. Replacement of IL-12 as an adjuvant by synthetic oligodeoxynucleotides (ODN) containing CpG motifs induced a similar level of vaccine-induced protection against challenge infection with P. chabaudi AS. These results illustrate that it is possible to enhance the potency of a crude malaria antigen preparation delivered in alum by inclusion of immunostimulatory molecules, such as IL-12 or CpG-ODN.

Retinoic acid and host-pathogen interactions: effects on inducible nitric oxide synthase in vivo

2000 ◽  
Vol 279 (5) ◽  
pp. E1045-E1053 ◽  
Author(s):  
Yvan Devaux ◽  
Sandrine Grosjean ◽  
Carole Seguin ◽  
Chantal David ◽  
Brigitte Dousset ◽  
...  
Keyword(s):  
Retinoic Acid ◽  
Interleukin 2 ◽  
In Vitro Study ◽  
Cell Types ◽  
Nitrite Accumulation ◽  
Ifn Γ ◽  
Wistar Kyoto ◽  
Plasma Nitrate

Vitamin A and its metabolite retinoic acid modulate the host response to pathogens through poorly characterized mechanisms. In vitro studies have suggested that retinoic acid decreases inducible NO synthase (NOS2, or iNOS) expression, a component of innate immunity, in several cell types stimulated with lipopolysaccharide (LPS) or cytokines. This study investigated the effect of retinoic acid on LPS-stimulated NOS2 expression in vivo. Wistar-Kyoto rats received all- trans retinoic acid (RA, 10 mg/kg) or vehicle intraperitoneally daily for 5 days followed by LPS (4 mg/kg) or saline intraperitoneally and were killed 6 h later. NOS2 activation was estimated by mRNA (RT-PCR) and protein (Western-blot) expression and plasma nitrate/nitrite accumulation. In sharp contrast to previous in vitro study reports, RA significantly enhanced NOS2 mRNA, protein expression, and plasma nitrate/nitrite concentration in LPS-injected rats but not in saline-injected rats. This was associated with increased expression of interleukin-2, interferon (IFN)-γ and IFN regulatory factor-1 mRNAs in several organs and increased IFN-γ plasma concentration. RA significantly increased mortality in LPS-injected rats. The NOS inhibitor aminoguanidine (50 mg/kg before LPS injection) significantly attenuated the RA-mediated increase in mortality. These results demonstrate for the first time that RA supplementation in vivo enhances activation of the LPS-triggered NOS2 pathway.

scholarly journals Mechanisms of T-Lymphocyte Accumulation during Experimental Pleural Infection Induced by Mycobacterium bovis BCG

2008 ◽  
Vol 76 (12) ◽  
pp. 5686-5693 ◽  
Author(s):  
Mariana C. Souza ◽  
Carmen Penido ◽  
Maria F. S. Costa ◽  
Maria Graças Henriques
Keyword(s):  
T Lymphocytes ◽  
Mycobacterium Bovis ◽  
T Lymphocyte ◽  
Mononuclear Cells ◽  
Interleukin 2 ◽  
Pleural Space ◽  
T Lymphocyte Proliferation ◽  
Pleural Infection ◽  
Ifn Γ

ABSTRACT Tuberculous pleurisy is a frequent extrapulmonary manifestation characterized by accumulation of fluid and inflammatory cells in the pleural space. Here, we investigated the mechanisms of T-lymphocyte accumulation in the pleural space by using a murine model of pleurisy induced by Mycobacterium bovis BCG. Intrathoracic (i.t.) injection of BCG (4.5 × 105 bacteria/cavity) induced accumulation of T lymphocytes in the pleural cavities of C57BL/6 mice. We observed the presence of CFU in pleural washes conducted 1, 2, 3, 7, and 15 days after pleurisy induction. Pretreatment with fucoidan inhibited T-lymphocyte accumulation at 1 day, but not at 15 days, after BCG-induced pleurisy. Accordingly, adoptive transfer of fluorescein isothiocyanate-labeled blood mononuclear cells to infected mice showed that T lymphocytes migrated into the pleural cavity 1 day (but not 15 days) after BCG injection. Cell-free pleural wash fluids recovered from mice 1 day after BCG i.t. stimulation (day 1 BCG-PW), but not day 7 or day 15 BCG-PW, induced in vitro T-cell transmigration, which was dependent on L-, P-, and E-selectins. In contrast, day 7 BCG-PW (but not day 1 BCG-PW) induced in vitro T-lymphocyte proliferation via interleukin-2 (IL-2) and gamma interferon (IFN-γ). Accordingly, in vivo IL-2 or IFN-γ neutralization abolished T-lymphocyte accumulation 7 days after pleurisy induction. Our results demonstrate that pleural infection induced by BCG leads to T-lymphocyte accumulation in two waves. The acute phase depends on selectin-mediated migration, while the second wave of T-lymphocyte accumulation seems to depend on a local proliferation induced by cytokines produced in situ.

scholarly journals An in vitro study to elucidate the effects of Septilin TM on immune pathways

2018 ◽  
Vol 17 (2) ◽  
pp. 238-244
Author(s):  
Mujeeb Hoosen ◽  
Edmund John Pool
Keyword(s):  
Medical Science ◽  
In Vitro Study ◽  
Cell Mediated Immunity ◽  
Immunomodulatory Effects ◽  
Ifn Γ ◽  
Herbal Medicinal ◽  
Immune Pathways ◽  
In Vitro Effects

Objective: The use of herbal immunomodulatory preparations to prevent and treat immunological complications is increasing in popularity. Rigorous in vitro, in vivo and clinical trial studies are needed to ensure safety, quality and efficacy for the wellbeing of patients. SeptilinTM, a proprietary herbal medicinal product has been reported to have immunomodulatory effects. This study investigated the in vitro effects of SeptilinTM on biomarkers of specific immune pathways by using whole blood culture assays (WBC).Materials and Methods: Stimulated and unstimulated WBC have been incubated with the product. Enzyme linked immunosorbent assays have been used to screen for IL-6, IL-10, and IFN γ as biomarkers for inflammation, humoral immunity, and cell mediated immunity, respectively.Results: SeptilinTM had no effect on the release of IL-6 production by (lipopolysaccharide) LPS stimulated WBC across all concentrations tested. However, SeptilinTM induced significant higher levels (P<0.001) of IL-6 release in unstimulated WBC across all concentrations between 0µg/ml-258µg/ml. SeptilinTM had no effect on the release of IL-10 release in unstimulated WBC across all concentrations. However the presence of SeptilinTM in phytohaemagglutinin (PHA) stimulated WBC induced significant higher release (P<0.01) of IL-10 release at concentrations between 64.5µg/ml- 258µg/ml when compared to the control. The presence of SeptilinTM in unstimulated WBC had no effect on the release of IFN γ production across all concentrations. The presence of SeptilinTM in PHA stimulated WBC release of IFN γ is inconclusive.Conclusion: This study shows that SeptilinTM has immunomodulatory effects on WBC in vitro.Bangladesh Journal of Medical Science Vol.17(2) 2018 p.238-244

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