ABSTRACT
The emergence of SARS-CoV-2, responsible for coronavirus disease-2019 (COVID-19), has become a major global health problem. The molecular testing is the accepted assay in SARS-CoV-2 detection. However, there are several reasons for low sensitivity by RNA detection, causing challenges in SARS-CoV-2 diagnosis. In this study, we aimed to investigate serological patterns of SARS-CoV-2 specific IgM, and IgG in 111 hospitalized, and 34 recovered COVID-19 patients and 311 prepandemic normal serum specimens by ELISA. The validity of the ELISA kits was evaluated using samples from normal and recovered cases. This showed that 98.1%, and 98.4% of prepandemic normal samples were negative for anti-SARS-CoV-2 IgM, and IgG, respectively. Assessment of 34 COVID-19 confirmed recovered patients showed a test sensitivity of 76.5%, and 94.1% for IgM, and IgG, respectively. In COVID-19 hospitalized patients, 42.3%, and 51.4% were positive for IgM and IgG, respectively. Viral RNA was not detectable in 43.3% of the hospitalized patients. Interestingly, combined molecular and serological testing improved the sensitivity of COVID-19 diagnosis to 79.6%. Using PCR with combined IgM/IgG results augmented the patient diagnosis sensitivity to 65.3% and 87.2% in ≤ 7 days, and > 7 days intervals, respectively. Overall, serological tests in combination with PCR can improve the sensitivity of COVID-19 diagnosis.
Subject(s)
Antibodies, Viral/blood , COVID-19 Serological Testing/methods , COVID-19/blood , COVID-19/diagnosis , Enzyme-Linked Immunosorbent Assay/methods , SARS-CoV-2/immunology , Adolescent , Adult , Aged , Aged, 80 and over , COVID-19/virology , Female , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Male , Middle Aged , Polymerase Chain Reaction , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification , Sensitivity and Specificity , Young AdultABSTRACT
[This corrects the article DOI: 10.1155/2021/5538348.].
ABSTRACT
An effective therapeutic vaccine to eradicate HIV-1 infection does not exist yet. Among different vaccination strategies, cell-based vaccines could achieve in clinical trials. Cell viability and low nucleic acid expression are the problems related to dendritic cells (DCs) and mesenchymal stem cells (MSCs), which are transfected with plasmid DNA. Thus, novel in vitro strategies are needed to improve DNA transfection into these cells. The recent study assessed immune responses generated by MSCs and DCs, which were derived from mouse bone marrow and modified with Nef antigen using novel methods in mice. For this purpose, an excellent gene transfection approach by mechanical methods was used. Our data revealed that the transfection efficacy of Nef DNA into the immature MSCs and DCs was improved by the combination of chemical and mechanical (causing equiaxial cyclic stretch) approaches. Also, chemical transfection performed two times with 48-hour intervals further increased gene expression in both cells. The groups immunized with Nef DC prime/rNef protein boost and then Nef MSC prime/rNef protein boost were able to stimulate high levels of IFN-γ, IgG2b, IgG2a, and Granzyme B directed toward Th1 responses in mice. Furthermore, the mesenchymal or dendritic cell-based immunizations were more effective compared to protein immunization for enhancement of the Nef-specific T-cell responses in mice. Hence, the use of chemical reagent and mechanical loading simultaneously can be an excellent method in delivering cargoes into DCs and MSCs. Moreover, DC- and MSC-based immunizations can be considered as promising approaches for protection against HIV-1 infections.
Subject(s)
AIDS Vaccines/immunology , HIV Infections/therapy , HIV-1/immunology , Transfection/methods , nef Gene Products, Human Immunodeficiency Virus/immunology , AIDS Vaccines/administration & dosage , AIDS Vaccines/genetics , Animals , Bioreactors , Dendritic Cells/immunology , Female , HIV Infections/immunology , HIV Infections/virology , HIV-1/genetics , Humans , Immunogenicity, Vaccine/genetics , Male , Mesenchymal Stem Cells/immunology , Mice , Models, Animal , Plasmids/genetics , Primary Cell Culture , Recombinant Proteins/administration & dosage , Recombinant Proteins/genetics , Recombinant Proteins/immunology , T-Lymphocytes/immunology , Transfection/instrumentation , nef Gene Products, Human Immunodeficiency Virus/administration & dosage , nef Gene Products, Human Immunodeficiency Virus/geneticsABSTRACT
INTRODUCTION: Long-term pulmonary complications are one of the major long-term consequences of sulfur mustard (SM) exposure. Toll-like receptor 4 (TLR4) involves in the pathogenesis of several pulmonary disorders. Surfactant protein-A (SP-A) regulates LPS-induced TLR4 localization and activation responses. However, the intensity and significance of TLR4 and SP-A expression by lung cells in SM-exposed patients is not clear. METHODS: The gene expression of TLR4 (through real-time PCR) and TLR4 and SP-A positive cells and alveolar type II cells, as SP-A producers, (using IHC) were assessed in formalin fixed paraffin embedded (FFPE) specimens from SM-exposed (nâ¯=â¯17), and non-SM exposed individuals (nâ¯=â¯12). RESULTS: TLR4 gene expression did not change between study groups. However, its cell surface presentation was significantly reduced in SM-exposed patients and particularly in which with constrictive bronchiolitis compared with the control group (Pâ¯<â¯0.001 and Pâ¯=â¯0.002, respectively). Frequency of alveolar type II cells was lower in the case group rather than the control group while the number of SP-A positive cells did not alter. CONCLUSIONS: These findings suggest that reduced TLR4 cell surface presentation may have anti-inflammatory function and SP-A may have a critical role in regulation of inflammatory responses in SM-exposed patients. Further investigation on other possible mechanisms involved in TLR4 internalization maybe help to illustrate the modulatory or inflammatory activity of TLR4 in these patients.
Subject(s)
Alveolar Epithelial Cells/pathology , Bronchiolitis Obliterans/chemically induced , Chemical Warfare Agents/toxicity , Mustard Gas/toxicity , Pulmonary Surfactant-Associated Protein A/metabolism , Toll-Like Receptor 4/metabolism , Adult , Alveolar Epithelial Cells/immunology , Bronchiolitis Obliterans/drug therapy , Bronchiolitis Obliterans/immunology , Bronchiolitis Obliterans/pathology , Case-Control Studies , Gene Expression Profiling , Humans , Immunohistochemistry , Iran , Male , Middle Aged , Pulmonary Surfactant-Associated Protein A/analysis , Time Factors , Toll-Like Receptor 4/analysisABSTRACT
OBJECTIVE: Dendritic cells (DCs) as major regulators of the immune response in the decidua play a pivotal role in establishment and maintenance of pregnancy. Immunological disorders are considered to be the main causes of unexplained recurrent spontaneous abortions (RSAs). Recently, we reported that mesenchymal stem cells (MSCs) therapy could improve fetal survival and reduce the abortion rate in abortion-prone mice, although the precise mechanisms of this action are poorly understood. Since MSCs have been shown to exert immunomodulatory effects on the immune cells, especially DCs, this study was performed to investigate the capability of MSCs to modulate the frequency, maturation state, and phenotype of uterine DCs (uDCs) as a potential mechanism for the improvement of pregnancy outcome. MATERIALS AND METHODS: In this experimental study, adipose-derived MSCs were intraperitoneally administered to abortion-prone pregnant mice on the fourth day of gestation. On the day 13.5 of pregnancy, after the determination of abortion rates, the frequency, phenotype, and maturation state of uDCs were analyzed using flow cytometry. RESULTS: Our results indicated that the administration of MSCs, at the implantation window, could significantly decrease the abortion rate and besides, increase the frequency of uDCs. MSCs administration also remarkably decreased the expression of DCs maturation markers (MHC-II, CD86, and CD40) on uDCs. However, we did not find any difference in the expression of CD11b on uDCs in MSCs-treated compared to control mice. CONCLUSION: Regarding the mutual role of uDCs in establishment of a particular immunological state required for appropriate implantation, proper maternal immune responses and development of successful pregnancy, it seems that the modulation of uDCs by MSCs could be considered as one of the main mechanisms responsible for the positive effect of MSCs on treatment of RSA.
ABSTRACT
Objective: Dendritic cells (DCs) are professional antigen presenting cells majorly modulated by various environmental factors. Leukemia inhibitory factor (LIF) is a pleiotropic cytokine from interleukin-6 family. Previous studies demonstrate that LIF is associated with several tolerogenic events; yet the exact effect of this cytokine on the generation and function of DCs was not explicitly identified. Materials and methods: To clarify the role of LIF in DCs development, immature DCs were differentiated from mouse bone marrow (BM) in a GM-CSF and IL-4 containing medium with or without LIF. Afterwards, in maturation process, the differentiated DCs were exposed to TNF-α in the presence or absence of LIF. Results: Immature DCs differentiated in the presence of LIF, proved a significant enhancement in the expression of MHCII, CD40, or CD86 molecules and in the antigen uptake function. LIF treatment of normal DCs while stimulating for maturation, caused a significant decrement in the expression of phenotypic markers as well as an increment in the antigen uptake function in comparison with TNF-α-only stimulated cells; however, the reduced ability for induction of allogenic T-cell proliferation proved no statistical significance. Conclusions: Our results can reflect a role for LIF in the generation and particularly maturation of DCs. It can be assumed that LIF rather modulates the maturation level, leading to the development of semi-mature and tolerogenic DCs. According to the high levels of LIF in immune-privileged sites like brain and uterine, it seems that the cytokine may account for the formation of local DCs that help the establishment of immunosuppressive environments.