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Exercise and Age-Related Decline in Immune Functions
Published in Ronald R. Watson, Marianne Eisinger, Exercise and Disease, 2020
Robert S. Mazzeo, Imran Nasrullah
Data from our laboratory have examined the effect of chronic endurance training on IL-2 synthesis, T-cell proliferation and cytotoxicity in Fischer 344 rats of 7, 17, and 27 months of age. The animals underwent 15 weeks of treadmill running at 75% maximal capacity, 5 days/week for 60 min. Controls remained sedentary throughout the period of the study, but were exposed to 5 min of running once per week. After the 15-week period, animals were sacrificed at least 24 h after their last exercise session. The spleens were removed aseptically, and splenocytes were collected, put into suspension, and used as stock for the various assays. Interleukin-2 was produced by exposing splenocytes to 5 μg/ml Con-A for 24 h; thereafter, supernatants were collected and passed through 0.22 μ filters. IL-2 produced was measured by the IL-2 dependent cell line CTLL-2 as an indicator population. T-cell proliferation was assessed by thymidine uptake upon exposure to varied doses of Con-A over a 48-h period. Natural cytotoxicity was measured using a chromium release assay with YAC-1 cell line serving as the target cell population for splenic effectors.
Perinatal exposure to silver nanoparticles reprograms immunometabolism and promotes pancreatic beta-cell death and kidney damage in mice
Published in Nanotoxicology, 2021
Ratnakar Tiwari, Radha Dutt Singh, Monika Binwal, Anurag Kumar Srivastav, Neha Singh, Hafizurrahman Khan, Siddhartha Gangopadhyay, Nidhi Argaria, Prem Narain Saxena, Somendu Kumar Roy, Mahadeo Kumar, Vineeta Sharma, Vikas Srivastava
To estimate the alterations in immune response, we performed splenocyte stimulation assays using isolated spleen cells from control and AgNPs exposed offspring. Splenocytes are a reservoir of blood cells, lymphocytes, and phagocytic cells. Stimulation with LPS and a mitogen in the assay gives an insight into prior immune conditioning of the host. Therefore, cultured splenocytes were exposed to LPS and ConA. Upon LPS treatment, splenocytes of the 0.5 ppm group showed increased cell proliferation (p < 0.05) but there was no significant change in proliferation in the 5.0 ppm group. Of note, the 50 ppm group showed reduced proliferation when compared to the LPS treated control group (Figure 4(A)). AgNPs exposed animals also showed alterations in the immune response against T cell mitogen ConA. While the 0.5 ppm group showed a comparable increase in proliferative response, the 5.0 ppm and 50 ppm groups showed significantly less proliferation as compared to ConA exposed control group (Figure 4(B)). To assess the immunological memory against AgNPs and Ag+ ions, splenocytes were exposed to various concentrations of AgNPs and AgNO3 (as the source of Ag+ ions). Notably, exposure to higher concentrations of AgNPs and Ag+ ions caused a reduction in proliferation in the 50 ppm group whereas the 0.5 and 5.0 ppm group did not show any significant change as compared to their respective treated controls (Figure 4(C,D)).
Current approaches toward identifying a correlate of immune protection from tuberculosis
Published in Expert Review of Vaccines, 2019
The majority of mouse studies utilize splenocytes for assessing vaccine-induced protection. Although splenocytes have been considered as strong predictors of vaccine efficacy, this is not translatable to human studies. Kurtz and Elkins developed an in vitro co-culture killing assay for testing the ability of murine immune peripheral blood lymphocytes, which would be more readily accessible for clinical studies to assess control of intracellular bacterial growth [160]. These studies incorporated gene expression profiling and protein expression assays using macrophages infected with BCG and M.tb in parallel. Immune peripheral blood lymphocytes from animals given a protective vaccine readily controlled intracellular infection. The authors highlighted that although these results support the value of this strategy as a functional correlate, such bioassays are difficult in clinical studies. The study revealed host-specific and mycobacterial-specific factors involved in protection against virulent M.tb. A set of 13 core molecules were induced in the most protected animals. The authors suggested that it is likely that any biosignature that correlates with vaccine efficacy against M.tb will be complex and multifactorial and advised on applying multivariate modeling [160].
Nicotine Augments the Beneficial Effects of Mesenchymal Stem Cell-based Therapy in Rat Model of Multiple Sclerosis
Published in Immunological Investigations, 2018
Shiva Khezri, Seyyed Meysam Abtahi Froushani, Mozhgan Shahmoradi
On day 33 after disease induction, the rats were euthanized under deep anesthesia and the spleens were aseptically removed from the rats. Next, the splenocytes were isolated by meshing the spleens through cell strainers (70 um) into a 50 mL conical tube. The splenocytes were prepared in RPMI-1640 medium, supplemented with 10% fetal calf serum, and the red blood cells were removed by RBC lysis buffer. The cell suspensions (2 × 106 cells/mL) were placed in 24-well plates and stimulated with PHA (10 ug/mL) (Abtahi Froushani et al., 2014; Abtahi Froushani, Esmaili Gouvarchin Galee et al. 2015). After 72 h, the culture supernatants were collected and checked by Enzyme-linked immunosorbent assay (ELISA) kits (for IFN-γ, IL-17, IL-10, and TNF-α), according to the manufacturer’s instructions (Abtahi Froushani et al., 2014, 2015).