Global, Regional, and National Prevalence of Overweight and Obesity in Children and Adults During 19802013: A Systematic Analysis for the Global Burden of Disease. Mech Ageing Dev (1998) 102:23. A Systems Biology Approach to Investigating the Influence of Exercise and Fitness on the Composition of Leukocytes in Peripheral Blood. J Immunol (2014) 193:12. doi:10.4049/jimmunol.1401685, 60. Regulation of Neutrophils in Type 2 Immune Responses. In addition, the expression of P16ink4a increases markedly with aging in most mouse tissues and in human skin and kidney tissues (45, 46), which suggests the importance of this tumor suppressor in aging and senescence (47, 48). Available at: https://cancerres.aacrjournals.org/content/60/8/2209. All samples were collected, processed, divided into aliquots, and stored at -80C for subsequent analysis. The patients/participants provided their written informed consent to participate in this study. Front Endocrinol (2019) 10:266. doi:10.3389/fendo.2019.00266, 32. Huttunem R, Syrjnem J. Obesity and the Risk and Outcome of Infection. J ImmunoTher Cancer (2019) 5:30. doi:10.1186/s40425-017-0231-8, 56. Junttila IS. Fulop T, Larbi A, Dupuis G, Le Page A, Frost EH, Cohen AA, et al. Sheridan PA, Paich HA, Handy J, Karlsson EA, Hudgens MG, Sammon AB, et al. Romagosa C, Simonetti S, Lpez-Vicente L, Mazo A, Lleonart ME, Castellvi J, et al. Obes (Silver Spring) (2016) 24:3. doi:10.1002/oby.21383, 14. As for the list of primers, the following 5 and 3 sequences were used: HPRT (forward: GACCAGTCAACAGGGGACAT, reverse: AACACTTCGTGGGGTCCTTTTC); Leptin (forward: GGCTTTGGTCCTATCTGTCTTATGTTC; reverse: CCTGTTGATAGACTGCCAGAGTCTG); CD27 (forward: ACTACTGGGCTCAGGGAAAGCT; reverse: GGATCACACTGAGCAGCCTTTC); CD28 (forward: GAGAAGAGCAATGGAACCATTATC; reverse: TAGCAAGCCAGGACTCCACCAA); P16INK4a (forward: GGGGGCACCAGAGGCAGT; reverse: GGTTGTGGCGGGGGCAGTT); PD1 (forward: CGTGGCCTATCCACTCCTCA; reverse: ATCCCTTGTCCCAGCCACTC); IL-2 (forward: GAATCCCAAACTCACCAGGATGCTC; reverse: TAGCACTTCCTCCAGAGGTTTGAGT); IL-6 (forward: GGTACATCCTCGACGGCATCT; reverse: GTGCCTCTTTGCTGCTTTCAC); IL-10 (forward: CCGTGGAGCAGGTGAAGAATG; reverse: AGTCGCCACCCTGATGTCTC); IL-4 forward: AACTTTGTCCACGGACACAAGTGC; reverse: GAATCGGATCAGCTGCTTGTGCCT); CCR7 (forward: GCCCAGATGGTTTTTGGGTTC; reverse: GCAAGGTACGGATGATAATGAGG); TNF (forward: CGAGTCTGGGCAGGTCTACTTT; reverse: AAGCTGTAGGCCCCAGTGAGTT). Williams A, Greene N, Kimbro K. Increased Circulating Cytokine Levels in African American Women With Obesity and Elevated HbA1c. Frasca D, Ferracci F, Diaz A, Romero M, Lechner S, Blomberg BB. (58) have reported an increase in the percentage of CD27+ B cells and NK cells during aging. Front Immunol (2018) 9:2741. doi:10.3389/fimmu.2018.02741, 37. Nat Med (2019) 25:14151. IUBMB Life (2007) 59:7. doi:10.1080/15216540701488358, 53. J Clin Invest (2014) 124:6. doi:10.1172/JCI72271, 33. The cardiorespiratory assessment was performed using a progressive effort protocol on a treadmill (Quinton, model TM55, USA), with a continuous collection of expired gases breath to breath (CPX Ultima, MedGraphics, USA). Cytotoxic Activity of CD4 T Cells During the Early Stage of Autoimmune Neuroinflammation. However, there is currently no complete understanding of the signs that trigger senescence and, although P16ink4a appears to be one of the main factors in senescence, more information is needed to determine the exact role of each factor in this process (54). Reidy K, Kang HM, Hostetter T, Susztak K. Molecular Mechanisms of Diabetic Kidney Disease. Nature (2001) 410:6824. doi:10.1038/35065016, 63. Front Immunol (2019) 10:2507. doi:10.3389/fimmu.2019.02507, 40. Obesity as a Model of Premature Immunosenescence. However, cell damage was seen to appear when these levels were increased (as in the case of obesity) and innate immunity cells, such as macrophages, exhibit an over-activated production of oxidative and inflammatory compounds which occurs mainly in the absence of antigenic stimulus and cannot be neutralized by antioxidant nor anti-inflammatory defenses, thus leading to a chronic state of oxidative and inflammatory stress (5, 10). Additionally, the extraction of the total leukocyte mRNA was performed following the instructions of the Trizol reagent. Immunol. Aging Cell (2006) 5:5. doi:10.1111/j.1474-9726.2006.00231.x, 47. Colonna-Romano G, Bulati M, Aquino A, Scialabba G, Candore G, Lio D, et al. Eur Heart J (2021) 42:34. doi:10.1093/eurheartj/ehab518, 4. The cardiorespiratory fitness verified by the maximum oxygen consumption (VO2max) was determined by the average values of the last 30 s of the test (29). Additionally, gene expression of IL-2 (p= 0.003 and p = 0.0001, respectively; Figure1B), IL4 (p = 0.001 and p = 0.011, respectively; Figure1C), CD27 (p = 0.014 and p = 0.043, respectively; Figure1D), IL10 (p = 0.001 and p = 0.0001, respectively; Figure1E), TNF- (p = 0.001 and p = 0.029,respectively; Figure1F), PD-1 (p = 0.006 and p = 0.003, respectively; Figure1G), P16ink4a (p = 0.014 and p = 0.029,respectively; Figure1J) and CCR7 (p = 0.0001 and p = 0.012, respectively; Figure1K) were significantly higher for OBD and OB compared to E, with no difference between OBD and OB for these same variables (p > 0.05 for all the comparisons). J Appl Physiol (1973) 35:2. doi:10.1152/jappl.1973.35.2.236, 30. Immunosenescence and Inflamm-Aging As Two Sides of the Same Coin: Friends or Foes? Diet-Induced Obesity Does Not Impact the Generation and Maintenance of Primary Memory CD8 T Cells. In addition, a significant correlation was found between VO2max and fat-free mass (r = .762, p = 0.0001). Sp1 is Essential for P16ink4a Expression in Human Diploid Fibroblasts During Senescence. Hunsche C, Hernandez O, de la Fuente M. Impaired Immune Response in Old Mice Suffering From Obesity and Premature Immunosenescence in Adulthood. Lancet (2014) 384:9945. doi:10.1016/S0140-6736(14)60460-8, 2. Additionally, the increase in adipose tissue and consequently in chronic inflammation and early senescence of the immune system can lead to impairments in the cardiorespiratory capacity of obese middle-aged individuals. Subjects were required to perform 10 repetitions at 50% of their estimated 1RM (according to each participants capacity). Akbar AN, Henson SM. DB and VB performed the gene expression analysis. A minimum of 15 ng cDNA per reaction was used for all genes in order to ensure the efficiency of the RT-PCR reaction. The association between physical fitness and body composition with the gene expression of the immunosenescence markers used in the present study was tested by Spearmans correlation test. Mol Cell Biol (1996) 16:3. doi:10.1128/mcb.16.3.859, 48. Peripheral Mononuclear Blood Cells Contribute to the Obesity-Associated Inflammatory State Independently of Glycemic Status: Involvement of the Novel Proinflammatory Adipokines Chemerin, Chitinase-3-Like Protein 1, Lipocalin-2 and Osteopontin. Several studies have proposed that the central mechanism underlying obesity and its related comorbidities comes from a persistent state of low-grade chronic inflammation together with dysregulation in the inflammation-stress feedback mechanisms (46), mainly due to increased production of pro-inflammatory markers such as leptin, interleukin-6 (IL-6), tumor necrosis factor - (TNF-), C-reactive protein (CRP) and monocyte chemotactic protein - 1 (MCP1) in the withe adipose tissue, together with the M1 macrophage phenotype infiltration in this tissue associated with obesity (68). 27. Diabetes (2015) 64:7. doi:10.2337/db14-1820, 26. Furthermore, no significant differences were observed in age, height and 1RM of the bench press and leg press between the groups studied (p > 0.05). J Lipid Res (2013) 54:9. doi:10.1194/jlr.M038638, 10. The Impact of Ageing on Natural Killer Cell Function and Potential Consequences for Health in Older Adults. *Correspondence: Diego T. Brunelli, diegotbrunelli@gmail.com, https://doi.org/10.3389/fimmu.2021.806400, https://www.asep.org/asep/asep/Brown2.pdf, https://cancerres.aacrjournals.org/content/60/8/2209, Creative Commons Attribution License (CC BY). Curr Pharm Des (2014) 20:29. doi:10.2174/1381612820666140130201734, 11. DB wrote the first draft of the manuscript. Furthermore, we observed that obesity could lead to impairments in cardiorespiratory fitness which has significant correlations with inflammation and the immunosenescence markers investigated in the present study. The editor and reviewer's affiliations are the latest provided on their Loop research profiles and may not reflect their situation at the time of review. Height was measured using a wall-mounted stadiometer with a precision of 0.1cm, and the weight was taken using a calibrated manual scale (Filizola S.A., So Paulo, SP, Brazil) with a precision of 0.1kg. Hellman J, Sansbury BE, Holden CR, Tang Y, Wong B, Wysoczynski M, et al. Waist circumference was measured at the midpoint between the last ribs and the iliac crests with an inelastic metric tape, with the precision of 0.1cm. All groups (OBD, OB and E) performed the functional analyses [muscle strength (1RM) and cardiorespiratory fitness (VO2max)], anthropometry, body composition (Air Displacement Plethysmograph), blood collections for biochemical (anti-CMV) and molecular (gene expression of leptin, IL-2, IL-4, IL-6, IL-10, TNF-, PD-1, P16ink4a, CCR7, CD28 and CD27) analyses of markers related to immunosenescence at baseline. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The measurement was performed in triplicate by the same trained professional and the average of these three measurements was calculated. Giordano A, Murano I, Mondini E, Perugini J, Smorlesi A, Severi I, et al. Cytokine Growth Factor Rev (2021) 59:9. doi:10.1016/j.cytogfr.2021.01.006, 19. However, the eutrophic individuals showed a tendency to express higher levels of CD28, in circulating leukocytes, than obese and diabetic individuals. CCR7 is expressed mainly in including nave T cells, central memory T cells, regulatory T cells, nave B cells, semi-mature/mature DCs and NK cells, and a minority of tumor cells, and it acts as a key regulator guiding homeostatic lymphocytes to secondary lymphoid organs (60). Furthermore, it has been proposed that dysfunctional adipocytes are involved in the production of oxidative stress through the generation of reactive oxygen species, implicating further inflammation and tissue damage (9). DB and RD performed the statistical analysis. Thus, after the initial evaluations, the middle-aged volunteers were allocated to each specific group: obese with T2D (OBD; n = 17; 7 males and 10 females); obese with absence of associated disease (OB; n = 18; 9 males and 9 females); and the eutrophic group (E; n = 18; 9 males and 9 females). DNA Damage Drives an Activin a-Dependent Induction of Cyclooxygenase-2 in Premalignant Cells and Lesions. PloS One (2007) 2:1. doi:10.1371/journal.pone.0000164, 50. Anti-CMV analysis was determined by enzyme-linked immunosorbent assay (ELISA), according to the specifications of the manufacturer (Novus Biologicals, catalog number KA1452, Abnova Corporation, USA). The BMI was calculated by dividing body mass (kg) by height squared (m). Canepa ET, Scassa ME, Ceruti JM, Marazita MC, Carcagno AL, Sirkin PF, et al. Table3 Correlations between physical fitness and body composition with gene expression of the immunosenescence markers of obese diabetic (OBD) or Obese without associated disease (OB) or Eutrophic (E) middle-aged individuals. It must be taken into consideration that this study is not without limitations. 29. Clin Mol Allergy (2017) 15:21. doi:10.1186/s12948-017-0077-0, 36. Diabetes (2016) 65:8. doi:10.2337/db15-1689, 64. Thus, we do not separate the results between CMV seropositive or negative individuals. Mech. For the production of the cDNA, we used the High Capacity cDNA Reverse Transcription Kit (Applied Biosystems, CA, USA), and followed the manufacturers instructions. Food records were given to the participants by trained researches who instructed them individually through a presentation of an already completed model food record and photographs of model home measures. Oncogene (2007) 26:55. doi:10.1038/sj.onc.1210578, 52. Figure1 Gene expression of leptin (A), IL-2 (B), IL-4 (C), CD27 (D), IL-10 (E), TNF- (F), PD-1 (G), CD28 (H), IL-6 (I), P16ink4a (J) and CCR7 (K) in PBMCs of obese diabetic (OBD) or Obese without associated disease (OB) or Eutrophic (E) middle-aged individuals. JEPonline (2001) 4:3. In animal models used in experimentation, Hunsche et al. Excess Body Mass Is Associated With T Cell Differentiation Indicative of Immune Ageing in Children. Given this context, the present study aimed to verify the effects of obesity per se or with associated comorbidity T2D on the gene expression of pro and anti-inflammatory and senescence markers of the immune system in circulating leukocytes and their correlations with physical fitness in obese middle-aged adults with T2D or without associated disease. Leptin expression was significantly higher for the OBD and OB groups compared to E (p = 0.001 and p= 0.047, respectively; Figure1A) and also for OBD compared to OB (p = 0.020; Figure1A). Ventura MT, Casciaro M, Gangemi S, Buquicchio R. Immunosenescence in Aging: Between Immune Cells Depletion and Cytokines Up-Regulation. Maurya R, Bhattacharya P, Dey R, Nakhasi HL. Additionally, all participants were informed about the purpose and risks of the study and signed an informed consent document before participation. In this sense, the present study aimed to verify, in circulating leukocytes, the gene expression of markers related to early immunosenescence associated with obesity and its possible relationships with the physical fitness in obese adults with type 2 diabetes or without associated comorbidities. Cataln V, Gmez-Ambrosi J, Rodrguez A, Ramrez B, Valent V, Moncada R, et al. doi:10.1038/nri2959, 44. Thus, since CD28 expression in our study trend to be lower in the obese groups, we speculate that chronic inflammation associated with obesity increases successive rounds of T cell proliferation and may induce exhaustion in memory CD8+ T cells, since these cells tend to lose CD28 expression before losing CD27 (57). The same investigator performed all measurement assessments. Thus, according to other studies that also found an increase in IL-4 associated with obesity (42), we speculate that increased expression of IL-4 in the obese groups compared to lean subjects may be due to a compensatory mechanism trying to maintain cell functions, homeostasis and tissue integrity. Front Immunol (2018) 8:1960. doi:10.3389/fimmu.2017.01960, 12. After 3min of rest, subjects were required to perform 3 repetitions at 70% of their estimated 1RM. This chronic inflammation associated with obesity directly impacts innate and adaptive immunity cells with both a pro-inflammatory orientation of immune cells in the basal state and results in a decrease in their ability to respond to infection and re-infection (6, 34). A Primer on Cytokines: Sources, Receptors, Effects, and Inducers. Oncogene (2011) 30:208797. Association Between HSV1 Seropositivity and Obesity: Data From the National Health and Nutritional Examination Surve2008. In this regard, since our results point to a negative correlation between the values of VO2max and CD27 expression, it is possible to infer that the persistence of CD27 + cells have some involvement in the context of dysregulation of the respiratory parameter in a systemic inflammatory process as observed in obesity. Ng M, Fleming T, Robinson M, Thomson B, Graetz N, Margono C, et al. Although the loss of the co-receptors CD27 and CD28 can be considered to be indicative of impaired telomere function in T cells and denotes progression towards replicative senescence (57), Colonna-Romano etal. Furthermore, we thank the volunteers who have participated in this study. Heeb LEM, Egholm C, Impellizzieri D, Ridder F, Boyman O. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.