Quantitation of Interferon γ mRNA Levels in Psoralen/UVA-Treated HUT-78 Cells by Competitive PCR

1994 ◽  
Vol 203 (2) ◽  
pp. 935-942 ◽  
Author(s):  
G.M. Saed ◽  
D.P. Fivenson
Keyword(s):  
Interferon Γ ◽  
Mrna Levels ◽  
Competitive Pcr

scholarly journals The usefulness of competitive PCR: airway gene expression of IL-5, IL-4, IL-4δ2, IL-2, and IFNγ in asthma

Thorax ◽  
2001 ◽  
Vol 56 (7) ◽  
pp. 541-548
Author(s):  
E M Glare ◽  
M Divjak ◽  
M J Bailey ◽  
E H Walters
Keyword(s):  
Gene Expression ◽  
Inhaled Corticosteroids ◽  
In Situ Hybridisation ◽  
Housekeeping Gene ◽  
Cross Sectional Study ◽  
Mrna Levels ◽  
Competitive Pcr ◽  
Cross Sectional ◽  
Steroid Use ◽  
Biopsy Specimens

BACKGROUNDAsthma has been described as an eosinophilic bronchitis driven by interleukin(IL)-4 and IL-5. The quantification of cytokine mRNA levels in airway samples has been confounded by housekeeping gene expression which differs between and within asthmatics and controls.METHODSThe usefulness of competitive reverse transcriptase-polymerase chain reaction (RT-PCR) that is independent of housekeeping gene expression for quantitating the mRNA for interferon (IFN)γ, IL-2, IL-5, IL-4 and its receptor antagonist encoding splicing variant IL-4δ2 was determined in a cross sectional study of 45 normal control subjects and 111 with asthma.RESULTSAtopic controls and atopic asthmatic subjects expressed more IL-5 than non-atopic controls (p<0.02) in bronchoalveolar lavage (BAL) cells, but not in biopsy specimens. IL-5 mRNA expression in BAL cells from asthmatic subjects using inhaled corticosteroids (ICS) was significantly lower than those not receiving ICS (p=0.04). IL-2 mRNA levels differed with steroid use in biopsy specimens but not in BAL cells. IFNγ, IL-4, and IL-4δ2 mRNA levels did not differ between any groups and were not affected by steroid use. IL-4 and IL-4δ2 mRNA levels were positively correlated (p<0.0001), suggesting coordinated transcription.CONCLUSIONSWhile the signal differentiation of competitive PCR in asthma may rival that of in situ hybridisation and immunohistochemistry, the method is expensive and wasteful of material.

scholarly journals Cell Culture-Derived Tilapia Lake Virus-Inactivated Vaccine Containing Montanide Adjuvant Provides High Protection against Viral Challenge for Tilapia

Vaccines ◽  
2021 ◽  
Vol 9 (2) ◽  
pp. 86
Author(s):  
Weiwei Zeng ◽  
Yingying Wang ◽  
Huzi Hu ◽  
Qing Wang ◽  
Sven M. Bergmann ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Protective Efficacy ◽  
Interferon Γ ◽  
Economic Losses ◽  
Mrna Levels ◽  
Infectious Dose ◽  
Booster Immunization ◽  
Virus Challenge ◽  
High Level ◽  
Factor Α

Tilapia lake virus (TiLV) is a newly emerging pathogen responsible for high mortality and economic losses in the global tilapia industry. Currently, no antiviral therapy or vaccines are available for the control of this disease. The goal of the present study was to evaluate the immunological effects and protective efficacy of formaldehyde- and β-propiolactone-inactivated vaccines against TiLV in the presence and absence of the Montanide IMS 1312 VG adjuvant in tilapia. We found that β-propiolactone inactivation of viral particles generated a vaccine with a higher protection efficacy against virus challenge than did formaldehyde. The relative percent survivals of vaccinated fish at doses of 108, 107, and 106 50% tissue culture infectious dose (TCID50)/mL were 42.9%, 28.5%, and 14.3% in the absence of the adjuvant and 85.7%, 64.3%, and 32.1% in its presence, respectively. The vaccine generated specific IgM and neutralizing antibodies against TiLV at 3 weeks following immunization that were significantly increased after a second booster immunization. The steady state mRNA levels of the genes tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interferon γ (IFN-γ), cluster of differentiation 4 (CD4), major histocompatibility complex (MHC)-Ia, and MHC-II were all increased and indicated successful immune stimulation against TiLV. The vaccine also significantly lowered the viral loads and resulted in significant increases in survival, indicating that the vaccine may also inhibit viral proliferation as well as stimulate a protective antibody response. The β-propiolactone-inactivated TiLV vaccine coupled with the adjuvant Montanide IMS 1312 VG and booster immunizations can provide a high level of protection from virus challenge in tilapia.

scholarly journals Angiogenesis-related factors in skeletal muscles of COPD patients: roles of angiopoietin-2

2013 ◽  
Vol 114 (9) ◽  
pp. 1309-1318 ◽  
Author(s):  
Mahroo Mofarrahi ◽  
Ioanna Sigala ◽  
Theodoros Vassilokopoulos ◽  
Sharon Harel ◽  
Yeting Guo ◽  
...  
Keyword(s):  
Vastus Lateralis ◽  
Interferon Γ ◽  
Forced Expiratory Volume ◽  
Chronic Obstructive ◽  
Mrna Levels ◽  
Gene Expressions ◽  
Related Factors ◽  
Control Subjects ◽  
Copd Patients ◽  
Angiopoietin 2

The role of angiogenesis factors in skeletal muscle dysfunction in patients with chronic obstructive pulmonary disease (COPD) is unknown. The first objective of this study was to assess various pro- and antiangiogenic factor and receptor expressions in the vastus lateralis muscles of control subjects and COPD patients. Preliminary inquiries revealed that angiopoietin-2 (ANGPT2) is overexpressed in limb muscles of COPD patients. ANGPT2 promotes skeletal satellite cell survival and differentiation. Factors that are involved in regulating muscle ANGPT2 production are unknown. The second objective of this study was to evaluate how oxidants and proinflammatory cytokines influence muscle-derived ANGPT2 expression. Angiogenic gene expressions in human vastus lateralis biopsies were quantified with low-density real-time PCR arrays. ANGPT2 mRNA expressions in cultured skeletal myoblasts were quantified in response to proinflammatory cytokine and H2O2 exposure. Ten proangiogenesis genes, including ANGPT2, were significantly upregulated in the vastus lateralis muscles of COPD patients. ANGPT2 mRNA levels correlated negatively with forced expiratory volume in 1 s and positively with muscle wasting. Immunoblotting confirmed that ANGPT2 protein levels were significantly greater in muscles of COPD patients compared with control subjects. ANGPT2 expression was induced by interferon-γ and -β and by hydrogen peroxide, but not by tumor necrosis factor. We conclude that upregulation of ANGPT2 expression in vastus lateralis muscles of COPD patients is likely due to oxidative stress and represents a positive adaptive response aimed at facilitating myogenesis and angiogenesis.

scholarly journals Synthetic Nano-selenium Improving Macrophage Immune Responses Treatment of Bladder Tumor Antigens

2021 ◽  
Vol 4 (1) ◽  
pp. 43-52
Author(s):  
Zeinab Agharezaie ◽  
◽  
Setareh Haghighat ◽  
Mohammad Hossein Yazdi ◽  
◽  
...  
Keyword(s):  
Bladder Tumor ◽  
Tumor Antigens ◽  
Mrna Level ◽  
Interleukin 10 ◽  
Interferon Γ ◽  
Selenium Nanoparticles ◽  
Maximum Effect ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Tumor Lysate

Background: Synthetic nanoparticles are deemed to improve treatment with the least adverse effects. The effect of Selenium Nanoparticles (SeNPs) was reviewed on various pathogenic disorders. In the present project, the role of SeNPs on macrophage responses was assessed. Materials and Methods: SeNPs were prepared synthetically by ascorbic acid. Macrophages (MQs) were cultured and treated with SeNPs in combination with bladder tumor lysate and Bacillus Calmette Guerin (BCG). Other experimental groups include SeNPs + tumor lysate + MQs, BCG, BCG+MQs, and MQs. The mRNA levels of interferon-γ and interleukin-10 were evaluated using the real-time PCR method. Results: Synthetic selenium nanoparticles combined with the tumor lysate upregulated the mRNA level of interferon-γ after 12 and 24 h treatment. Regarding interleukin-10 expression, there were no remarkable differences in all experimental groups. The maximum effect of synthetic SeNPs was observed after 24 h treatment. Conclusion: The optimum effect of synthetic SeNPs presents in a treatment-dependent manner.

scholarly journals Reproducibility in the Quantification of mRNA Levels by RT-PCR-ELISA and RT Competitive-PCR-ELISA

BioTechniques ◽  
1998 ◽  
Vol 24 (4) ◽  
pp. 652-658 ◽  
Author(s):  
L.L. Hall ◽  
G.R. Bicknell ◽  
L. Primrose ◽  
J.H. Pringle ◽  
J.A. Shaw ◽  
...  
Keyword(s):  
Mrna Levels ◽  
Competitive Pcr ◽  
Rt Pcr

scholarly journals Histological and immunological differences between zoonotic cutaneous leishmaniasis due to Leishmania major and sporadic cutaneous leishmaniasis due to Leishmania infantum

Parasite ◽  
2019 ◽  
Vol 26 ◽  
pp. 9 ◽  
Author(s):  
Thouraya Boussoffara ◽  
Mohamed Samir Boubaker ◽  
Melika Ben Ahmed ◽  
Mourad Mokni ◽  
Ikram Guizani ◽  
...  
Keyword(s):  
Cutaneous Leishmaniasis ◽  
Leishmania Major ◽  
Histological Analysis ◽  
Quantitative Polymerase Chain Reaction ◽  
Immunohistochemical Analysis ◽  
Interferon Γ ◽  
Mrna Levels ◽  
Zoonotic Cutaneous Leishmaniasis ◽  
Chemotactic Protein ◽  
Polymerase Chain

Lesion features in cutaneous leishmaniasis (CL) depend on the infecting Leishmania species as well as on host immune reponse. In this study, we evaluated the histological and immunological differences between two forms of CL described in Tunisia: zoonotic cutaneous leishmaniasis (ZCL) caused by L. major and sporadic cutaneous leishmaniasis (SCL) caused by L. infantum. Histological analysis showed a mild to moderate infiltrate within ZCL lesions. In contrast, massive infiltration of the dermis was observed within SCL lesions. Contrary to ZCL, infiltrates within SCL lesions were organized and showed granuloma composed of macrophages and lymphocytes. In addition, immunohistochemical analysis showed a predominance of CD4+ T cells within both CL forms. Furthermore, expression of interferon-γ, interleukin (IL)-10, IL-8, IL-13 and monocyte chemotactic protein (MCP)-1 was evaluated using real-time quantitative polymerase chain reaction (RT-qPCR). MCP-1 and IL-10 were expressed at comparable levels in ZCL and SCL lesions. Interestingly, IL-8 mRNA levels were significantly higher in ZCL lesions compared to SCL lesions, but interferon-γ was significantly higher in SCL lesions than in ZCL lesions.

Stimulation of toll-like receptor 4 expression in human mononuclear phagocytes by interferon-γ: a molecular basis for priming and synergism with bacterial lipopolysaccharide

Blood ◽  
2002 ◽  
Vol 99 (9) ◽  
pp. 3427-3431 ◽  
Author(s):  
Daniela Bosisio ◽  
Nadia Polentarutti ◽  
Marina Sironi ◽  
Sergio Bernasconi ◽  
Kensuke Miyake ◽  
...  
Keyword(s):  
Surface Expression ◽  
Interferon Γ ◽  
Mononuclear Phagocytes ◽  
Bacterial Lipopolysaccharide ◽  
Interleukin 12 ◽  
Mrna Levels ◽  
Adapter Protein ◽  
Toll Like Receptor ◽  
Toll Like Receptor 4 ◽  
Tlr4 Mrna

Abstract In human monocytes and macrophages, interferon-γ (IFNγ) augmented mRNA and surface expression of toll-like receptor 4 (TLR4), a crucial component of the signaling receptor complex for bacterial lipopolysaccharide (LPS). Expression of the accessory component MD-2 and of the adapter protein MyD88 was also increased. LPS increased TLR4 mRNA levels, but concomitantly decreased its surface expression. IFNγ counteracted the LPS-induced downregulation of TLR4. IFNγ-primed monocytes showed increased responsiveness to LPS in terms of phosphorylation of the interleukin-1 receptor–associated kinase (IRAK; immediately downstream of the MyD88 adapter protein), NF-kB DNA binding activity, and, accordingly, of cytokine (tumor necrosis factor α [TNFα] and interleukin-12 [IL-12]) production. These results suggest that enhanced TLR4 expression underlies the long-known priming by IFNγ of mononuclear phagocytes for pathogen recognition and killing as well as its synergism with LPS in macrophage activation.

scholarly journals Gonadotropin-Releasing Hormone in Regulation of Thymic Development in Rats: Profile of Thymic Cytokines

2019 ◽  
Vol 20 (16) ◽  
pp. 4033 ◽  
Author(s):  
Victoria I. Melnikova ◽  
Nadezhda V. Lifantseva ◽  
Svetlana N. Voronova ◽  
Liudmila A. Zakharova
Keyword(s):  
Ex Vivo ◽  
Interferon Γ ◽  
Gonadotropin Releasing Hormone ◽  
Recent Experimental Data ◽  
Mrna Levels ◽  
Fetal Thymus ◽  
Releasing Hormone ◽  
Thymus Development ◽  
Ex Vivo Culture ◽  
The Impact

An increasing body of recent experimental data confirms the impact of neurohormones on fetal development and function of different body systems. The synthesis of many neurohormones starts in fetal tissues before the hypothalamic–pituitary–adrenal and hypothalamic–pituitary–gonadal systems are formed, and their high levels are detected in the bloodstream. Here, we studied the role of gonadotropin-releasing hormone (GnRH) in rat thymus development and tried to reveal possible mechanisms underlying the GnRH effects in early development. Western blotting and reverse transcription-polymerase chain reaction allowed us to identify receptor for GnRH in the fetal thymus with peak expression on embryonic days 17–18 (ED17–18). Blocking the receptors in utero on ED17 by a GnRH antagonist suppressed the concanavalin A-induced proliferative response of T cells in adults. GnRH (10−7 M) increased mRNA expression of interleukin (IL)-4, IL-10, IL-1β, interferon γ (IFNγ), and tumor necrosis factor α (TNFα) in the thymus of 18-day fetuses after an ex vivo culture for 24 h. The increased mRNA levels of the cytokines in the thymus were accompanied by increased numbers of CD4+ T helpers. Overall, the data obtained confirm the regulatory or morphogenetic effect of GnRH on fetal thymus development mediated by synthesis of thymic cytokines.

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