Biological sex, not reproductive cycle, influences peripheral blood immune cell prevalence in mice.
blood
flow cytometry
immunology
immunophenotype
reproductive cycle
sex differences
Journal
The Journal of physiology
ISSN: 1469-7793
Titre abrégé: J Physiol
Pays: England
ID NLM: 0266262
Informations de publication
Date de publication:
04 2021
04 2021
Historique:
received:
14
08
2020
accepted:
03
12
2020
pubmed:
19
1
2021
medline:
21
4
2021
entrez:
18
1
2021
Statut:
ppublish
Résumé
Traditionally the female sex, compared with the male sex, has been perceived as having greater variability in many physiological traits, including within the immune system. We investigated effects of biological sex and the female reproductive cycle on numbers of circulating leukocytes in C57BL/6J mice. We show that biological sex, but not female reproductive cyclicity, has a significant effect on peripheral blood immune cell prevalence and variability, and that sex differences were not consistent amongst common inbred laboratory mouse strains. We found that male C57BL/6J mice, compared with female mice, have greater variability in peripheral blood immunophenotype, and that this was influenced by body weight. We created summary tables for researchers to facilitate experiment planning and sample size calculations for peripheral immune cells that consider the effects of biological sex. Immunophenotyping (i.e. quantifying the number and types of circulating leukocytes) is used to characterize immune changes during health and disease, and in response to pharmacological and other interventions. Despite the importance of biological sex in immune function, there is considerable uncertainty amongst researchers as to the extent to which biological sex or the female reproductive cycle influence blood immunophenotype. We quantified circulating leukocytes by multicolour flow cytometry in young C57BL/6J mice and assessed the effects of the reproductive cycle, biological sex, and other experimental and biological factors on data variability. We found that there are no significant effects of the female reproductive cycle on the prevalence of peripheral blood B cells, NK cells, CD4
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2169-2195Subventions
Organisme : CIHR
Pays : Canada
Commentaires et corrections
Type : CommentIn
Informations de copyright
© 2021 The Authors. The Journal of Physiology © 2021 The Physiological Society.
Références
Bain BJ & England JM (1975). Variations in leucocyte count during menstrual cycle. Br Med J 2, 473-475.
Baker M (2016). 1,500 scientists lift the lid on reproducibility. Nature 533, 452-454.
Bartke A & Dalterio S (1975). Evidence for episodic secretion of testosterone in laboratory mice. Steroids 26, 749-756.
Basson AR, LaSalla A, Lam G, Kulpins D, Moen EL, Sundrud MS, Miyoshi J, Ilic S, Theriault BR, Cominelli F & Rodriguez-Palacios A (2020). Artificial microbiome heterogeneity spurs six practical action themes and examples to increase study power-driven reproducibility. Sci Rep 10, 5039.
Becker JB, Arnold AP, Berkley KJ, Blaustein JD, Eckel LA, Hampson E, Herman JP, Marts S, Sadee W, Steiner M, Taylor J & Young E (2005). Strategies and Methods for Research on Sex Differences in Brain and Behavior. Endocrinology 146, 1650-1673.
Becker JB, Prendergast BJ & Liang JW (2016). Female rats are not more variable than male rats: a meta-analysis of neuroscience studies. Biol Sex Differ 7, 34.
Beery AK (2018). Inclusion of females does not increase variability in rodent research studies. Curr Opin Behav Sci 23, 143-149.
Beery AK & Zucker I (2011). Sex bias in neuroscience and biomedical research. Neurosci Biobehav Rev 35, 565-572.
Bogue MA, Grubb SC, Walton DO, Philip VM, Kolishovski G, Stearns T, Dunn MH, Skelly DA, Kadakkuzha B, TeHennepe G, Kunde-Ramamoorthy G & Chesler EJ (2018). Mouse Phenome Database: an integrative database and analysis suite for curated empirical phenotype data from laboratory mice. Nucleic Acids Res 46, D843-D850.
Bouman A, Moes H, Heineman MJ, de Leij LF & Faas MM (2001a). Cytokine production by natural killer lymphocytes in follicular and luteal phase of the ovarian cycle in humans. Am J Reprod Immunol 45, 130-134.
Bouman A, Moes H, Heineman MJ, de Leij LF & Faas MM (2001b). The immune response during the luteal phase of the ovarian cycle: increasing sensitivity of human monocytes to endotoxin. Fertil Steril 76, 555-559.
Brodin P & Davis MM (2017). Human immune system variation. Nat Rev Immunol 17, 21-29.
Brodin P, Jojic V, Gao T, Bhattacharya S, Angel CJ, Furman D, Shen-Orr S, Dekker CL, Swan GE, Butte AJ, Maecker HT & Davis MM (2015). Variation in the human immune system is largely driven by non-heritable influences. Cell 160, 37-47.
Buch T, Moos K, Ferreira FM, Fröhlich H, Gebhard C & Tresch A (2019). Benefits of a factorial design focusing on inclusion of female and male animals in one experiment. J Mol Med 97, 871-877.
Burden N, Chapman K, Sewell F & Robinson V (2015). Pioneering better science through the 3Rs: an introduction to the national centre for the replacement, refinement, and reduction of animals in research (NC3Rs). J Am Assoc Lab Anim Sci 54, 198-208.
Byers SL, Wiles MV, Dunn SL & Taft RA (2012). Mouse estrous cycle identification tool and images. PLoS One 7, e35538.
Carr EJ, Dooley J, Garcia-Perez JE, Lagou V, Lee JC, Wouters C, Meyts I, Goris A, Boeckxstaens G, Linterman MA & Liston A (2016). The cellular composition of the human immune system is shaped by age and cohabitation. Nat Immunol 17, 461-468.
Celec P, Ostatníková D & Hodosy J (2015). On the effects of testosterone on brain behavioral functions. Front Neurosci 9, 12-12.
Chan KA, Jazwiec PA, Gohir W, Petrik JJ & Sloboda DM (2018). Maternal nutrient restriction impairs young adult offspring ovarian signaling resulting in reproductive dysfunction and follicle loss. Biol Reprod 98, 664-682.
Check Hayden E (2010). Sex bias blights drug studies. Nature 464, 332-333.
Chuang KH, Altuwaijri S, Li G, Lai JJ, Chu CY, Lai KP, Lin HY, Hsu JW, Keng P, Wu MC & Chang C (2009). Neutropenia with impaired host defense against microbial infection in mice lacking androgen receptor. J Exp Med 206, 1181-1199.
Clayton JA (2016). Studying both sexes: a guiding principle for biomedicine. FASEB J 30, 519-524.
Clayton JA & Collins FS (2014). Policy: NIH to balance sex in cell and animal studies. Nature 509, 282-283.
Collins FS & Tabak LA (2014). Policy: NIH plans to enhance reproducibility. Nature 505, 612-613.
Coquelin A & Desjardins C (1982). Luteinizing hormone and testosterone secretion in young and old male mice. Am J Physiol 243, E257-263.
Cora MC, Kooistra L & Travlos G (2015). Vaginal Cytology of the laboratory rat and mouse: review and criteria for the staging of the estrous cycle using stained vaginal smears. Toxicol Pathol 43, 776-793.
Correa-De-Araujo R (2006). Serious gaps: how the lack of sex/gender-based research impairs health. J Womens Health 15, 1116-1122.
Darmochwal-Kolarz D, Rolinski J, Tabarkiewicz J, Leszczynska-Gorzelak B, Buczkowski J, Wojas K & Oleszczuk J (2003). Blood myeloid and lymphoid dendritic cells are stable during the menstrual cycle but deficient during mid-gestation. J Reprod Immunol 59, 193-203.
Dayton A, Exner EC, Bukowy JD, Stodola TJ, Kurth T, Skelton M, Greene AS & Cowley AW, Jr (2016). Breaking the cycle: estrous variation does not require increased sample size in the study of female rats. Hypertension 68, 1139-1144.
De Gregorio PR, Salva S, Tomás MST & Nader-Macias MEF (2018). Effects of exogenous sex hormones on mouse estrous cycle, vaginal microbiota and immune cells. Scand J Lab Anim Sci 44, 14.
Demas GE & Carlton ED (2015). Ecoimmunology for psychoneuroimmunologists: Considering context in neuroendocrine-immune-behavior interactions. Brain Behav Immun 44, 9-16.
Diaz Brinton R (2012). Minireview: translational animal models of human menopause: challenges and emerging opportunities. Endocrinology 153, 3571-3578.
Faas M, Bouman A, Moesa H, Heineman MJ, de Leij L & Schuiling G (2000). The immune response during the luteal phase of the ovarian cycle: a Th2-type response?. Fertil Steril 74, 1008-1013.
Faul F, Erdfelder E, Lang AG & Buchner A (2007). G*Power 3: a flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behav Res Methods 39, 175-191.
Festing MFW & Altman DG (2002). Guidelines for the design and statistical analysis of experiments using laboratory animals. ILAR J 43, 244-258.
Fields RD (2014). NIH policy: Mandate goes too far. Nature 510, 340-340.
Fish EN (2008). The X-files in immunity: sex-based differences predispose immune responses. Nat Rev Immunol 8, 737-744.
Fleiss JL & Cohen J (1973). The equivalence of weighted kappa and the intraclass correlation coefficient as measures of reliability. Educ Psychol Meas 33, 613-619.
Galea LAM, Choleris E, Albert AYK, McCarthy MM & Sohrabji F (2020). The promises and pitfalls of sex difference research. Front Neuroendocrinol 56, 100817.
Geller SE, Koch AR, Roesch P, Filut A, Hallgren E & Carnes M (2018). The more things change, the more they stay the same: A study to evaluate compliance with inclusion and assessment of women and minorities in randomized controlled trials. Acad Med 93, 630-635.
Gloskowska-Moraczewska Z & Sitarska E (1973). [Blood pattern in white rats in the course of sexual cycle]. Acta Haematol Pol 4, 117-121.
Gobinath AR, Choleris E & Galea LA (2017). Sex, hormones, and genotype interact to influence psychiatric disease, treatment, and behavioral research. J Neurosci Res 95, 50-64.
Grundy D (2015). Principles and standards for reporting animal experiments in the journal of physiology and experimental physiology. J Physiol 593, 2547-2549.
Halsey LG, Curran-Everett D, Vowler SL & Drummond GB (2015). The fickle P value generates irreproducible results. Nat Methods 12, 179-185.
Higginson AD & Munafo MR (2016). Current incentives for scientists lead to underpowered studies with erroneous conclusions. PLoS Biol 14, e2000995.
Hoggatt J, Hoggatt AF, Tate TA, Fortman J & Pelus LM (2016). Bleeding the laboratory mouse: Not all methods are equal. Exp Hematol 44, 132-137.e1.e131.
Horii Y, Nagasawa T, Sakakibara H, Takahashi A, Tanave A, Matsumoto Y, Nagayama H, Yoshimi K, Yasuda MT, Shimoi K & Koide T (2017). Hierarchy in the home cage affects behaviour and gene expression in group-housed C57BL/6 male mice. Sci Rep 7, 6991.
Ingvorsen C, Karp NA & Lelliott CJ (2017). The role of sex and body weight on the metabolic effects of high-fat diet in C57BL/6N mice. Nutr Diabetes 7, e261-e261.
Itoh Y & Arnold AP (2015). Are females more variable than males in gene expression? Meta-analysis of microarray datasets. Biol Sex Differ 6, 18.
Jasienska G, Bribiescas RG, Furberg A-S, Helle S & Núñez-de la Mora A (2017). Human reproduction and health: an evolutionary perspective. Lancet North Am Ed 390, 510-520.
Kaczorowski KJ, Shekhar K, Nkulikiyimfura D, Dekker CL, Maecker H, Davis MM, Chakraborty AK & Brodin P (2017). Continuous immunotypes describe human immune variation and predict diverse responses. Proc Natl Acad Sci 114, E6097.
Kappel S, Hawkins P & Mendl MT (2017). To group or not to group? Good practice for housing male laboratory mice. Animals 7, 88.
Karp NA, Mason J, Beaudet AL, Benjamini Y, Bower L, Braun RE, Brown SDM, Chesler EJ, Dickinson ME, Flenniken AM, Fuchs H, Angelis MHd, Gao X, Guo S, Greenaway S, Heller R, Herault Y, Justice MJ, Kurbatova N, Lelliott CJ, Lloyd KCK, Mallon A-M, Mank JE, Masuya H, McKerlie C, Meehan TF, Mott RF, Murray SA, Parkinson H, Ramirez-Solis R, Santos L, Seavitt JR, Smedley D, Sorg T, Speak AO, Steel KP, Svenson KL, Obata Y, Suzuki T, Tamura M, Kaneda H, Furuse T, Kobayashi K, Miura I, Yamada I, Tanaka N, Yoshiki A, Ayabe S, Clary DA, Tolentino HA, Schuchbauer MA, Tolentino T, Aprile JA, Pedroia SM, Kelsey L, Vukobradovic I, Berberovic Z, Owen C, Qu D, Guo R, Newbigging S, Morikawa L, Law N, Shang X, Feugas P, Wang Y, Eskandarian M, Zhu Y, Nutter LMJ, Penton P, Laurin V, Clarke S, Lan Q, Sohel K, Miller D, Clark G, Hunter J, Cabezas J, Bubshait M, Carroll T, Tondat S, MacMaster S, Pereira M, Gertsenstein M, Danisment O, Jacob E, Creighton A, Sleep G, Clark J, Teboul L, Fray M, Caulder A, Loeffler J, Codner G, Cleak J, Johnson S, Szoke-Kovacs Z, Radage A, Maritati M, Mianne J, Gardiner W, Allen S, Cater H, Stewart M, Keskivali-Bond P, Sinclair C, Brown E, Doe B, Wardle-Jones H, Grau E, Griggs N, Woods M, Kundi H, Griffiths MND, Kipp C, Melvin DG, Raj NPS, Holroyd SA, Gannon DJ, Alcantara R, Galli A, Hooks YE, Tudor CL, Green AL, Kussy FL, Tuck EJ, Siragher EJ, Maguire SA, Lafont DT, Vancollie VE, Pearson SA, Gates AS, Sanderson M, Shannon C, Anthony LFE, Sumowski MT, McLaren RSB, Swiatkowska A, Isherwood CM, Cambridge EL, Wilson HM, Caetano SS, Mazzeo CI, Dabrowska MH, Lillistone C, Estabel J, Maguire AKB, Roberson L-A, Pavlovic G, Birling M-C, Marie W-D, Jacquot S, Ayadi A, Ali-Hadji D, Charles P, André P, Le Marchand E, El Amri A, Vasseur L, Aguilar-Pimentel A, Becker L, Treise I, Moreth K, Stoeger T, Amarie OV, Neff F, Wurst W, Bekeredjian R, Ollert M, Klopstock T, Calzada-Wack J, Marschall S, Brommage R, Steinkamp R, Lengger C, Östereicher MA, Maier H, Stoeger C, Leuchtenberger S, Yildrim A, Garrett L, Hölter SM, Zimprich A, Seisenberger C, Bürger A, Graw J, Eickelberg O, Zimmer A, Wolf E, Busch DH, Klingenspor M, Schmidt-Weber C, Gailus-Durner V, Beckers J, Rathkolb B, Rozman J, Wakana S, West D, Wells S, Westerberg H, Yaacoby S, White JK & International Mouse Phenotyping C (2017). Prevalence of sexual dimorphism in mammalian phenotypic traits. Nat Commun 8, 15475.
Kilkenny C, Parsons N, Kadyszewski E, Festing MF, Cuthill IC, Fry D, Hutton J & Altman DG (2009). Survey of the quality of experimental design, statistical analysis and reporting of research using animals. PLoS One 4, e7824.
Klein SL, Schiebinger L, Stefanick ML, Cahill L, Danska J, de Vries GJ, Kibbe MR, McCarthy MM, Mogil JS, Woodruff TK & Zucker I (2015). Opinion: Sex inclusion in basic research drives discovery. Proc Natl Acad Sci 112, 5257.
Lee S, Kim J, Jang B, Hur S, Jung U, Kil K, Na B, Lee M, Choi Y, Fukui A, Gilman-Sachs A & Kwak-Kim JY (2010). Fluctuation of peripheral blood T, B, and NK cells during a menstrual cycle of normal healthy women. J Immunol 185, 756.
Machida T, Yonezawa Y & Noumura T (1981). Age-associated changes in plasma testosterone levels in male mice and their relation to social dominance or subordinance. Horm Behav 15, 238-245.
Mangino M, Roederer M, Beddall MH, Nestle FO & Spector TD (2017). Innate and adaptive immune traits are differentially affected by genetic and environmental factors. Nat Commun 8, 13850.
Mathur S, Mathur RS, Goust JM, Williamson HO & Fudenberg HH (1979). Cyclic variations in white cell subpopulations in the human menstrual cycle: correlations with progesterone and estradiol. Clin Immunol Immunopathol 13, 246-253.
McEwen BS, Gray JD & Nasca C (2015). 60 YEARS OF NEUROENDOCRINOLOGY: Redefining neuroendocrinology: stress, sex and cognitive and emotional regulation. J Endocrinol 226, T67-T83.
Moen EL, Fricano-Kugler CJ, Luikart BW & O'Malley AJ (2016). Analyzing clustered data: why and how to account for multiple observations nested within a study participant?. PLoS One 11, e0146721-e0146721.
Mogil JS & Chanda ML (2005). The case for the inclusion of female subjects in basic science studies of pain. Pain 117, 1-5.
Mysliwska J, Stech J, Skassa W & Mysliwski A (1975). [Peripheral blood lymphocytes in female mice during successive phases of the sexual cycle]. Acta Haematol Pol 6, 37-43.
Nakagawa S & Cuthill IC (2007). Effect size, confidence interval and statistical significance: a practical guide for biologists. Biol Rev Camb Philos Soc 82, 591-605.
Northern AL, Rutter SM & Peterson CM (1994). Cyclic changes in the concentrations of peripheral blood immune cells during the normal menstrual cycle. Proc Soc Exp Biol Med 207, 81-88.
Nowak J, Borkowska B & Pawlowski B (2016). Leukocyte changes across menstruation, ovulation, and mid-luteal phase and association with sex hormone variation. Am J Hum Biol 28, 721-728.
Nyby JG (2008). Reflexive testosterone release: a model system for studying the nongenomic effects of testosterone upon male behavior. Front Neuroendocrinol 29, 199-210.
Oellrich A, Meehan TF, Parkinson H, Sarntivijai S, White JK & Karp NA (2016). Reporting phenotypes in mouse models when considering body size as a potential confounder. J Biomed Semantics 7, 2.
Orru V, Steri M, Sole G, Sidore C, Virdis F, Dei M, Lai S, Zoledziewska M, Busonero F, Mulas A, Floris M, Mentzen WI, Urru SA, Olla S, Marongiu M, Piras MG, Lobina M, Maschio A, Pitzalis M, Urru MF, Marcelli M, Cusano R, Deidda F, Serra V, Oppo M, Pilu R, Reinier F, Berutti R, Pireddu L, Zara I, Porcu E, Kwong A, Brennan C, Tarrier B, Lyons R, Kang HM, Uzzau S, Atzeni R, Valentini M, Firinu D, Leoni L, Rotta G, Naitza S, Angius A, Congia M, Whalen MB, Jones CM, Schlessinger D, Abecasis GR, Fiorillo E, Sanna S & Cucca F (2013). Genetic variants regulating immune cell levels in health and disease. Cell 155, 242-256.
Parker RM & Browne WJ (2014). The place of experimental design and statistics in the 3Rs. ILAR J 55, 477-485.
Pennell LM, Galligan CL & Fish EN (2012). Sex affects immunity. J Autoimmun 38, J282-J291.
Pick R, He W, Chen C-S & Scheiermann C (2019). Time-of-day-dependent trafficking and function of leukocyte subsets. Trends Immunol 40, 524-537.
Piper SK, Grittner U, Rex A, Riedel N, Fischer F, Nadon R, Siegerink B & Dirnagl U (2019). Exact replication: Foundation of science or game of chance? PLoS Biol 17, e3000188.
Prasad A, Mumford SL, Buck Louis GM, Ahrens KA, Sjaarda LA, Schliep KC, Perkins NJ, Kissell KA, Wactawski-Wende J & Schisterman EF (2014). Sexual activity, endogenous reproductive hormones and ovulation in premenopausal women. Horm Behav 66, 330-338.
Prendergast BJ, Onishi KG & Zucker I (2014). Female mice liberated for inclusion in neuroscience and biomedical research. Neurosci Biobehav Rev 40, 1-5.
Ritz SA, Antle DM, Cote J, Deroy K, Fraleigh N, Messing K, Parent L, St-Pierre J, Vaillancourt C & Mergler D (2014). First steps for integrating sex and gender considerations into basic experimental biomedical research. FASEB J 28, 4-13.
Shansky RM (2019). Are hormones a “female problem” for animal research? Science 364, 825.
Shaw R, Festing MF, Peers I & Furlong L (2002). Use of factorial designs to optimize animal experiments and reduce animal use. ILAR J 43, 223-232.
Shen-Orr SS, Furman D, Kidd BA, Hadad F, Lovelace P, Huang YW, Rosenberg-Hasson Y, Mackey S, Grisar FA, Pickman Y, Maecker HT, Chien YH, Dekker CL, Wu JC, Butte AJ & Davis MM (2016). Defective Signaling in the JAK-STAT Pathway Tracks with Chronic Inflammation and Cardiovascular Risk in Aging Humans. Cell Syst 3, 374-384.e4.e374.
Smith AJ, Clutton RE, Lilley E, Hansen KEA & Brattelid T (2018). PREPARE: guidelines for planning animal research and testing. Lab Anim 52, 135-141.
Tannenbaum C, Schwarz JM, Clayton JA, de Vries GJ & Sullivan C (2016). Evaluating sex as a biological variable in preclinical research: the devil in the details. Biol Sex Differ 7, 13.
ter Horst JP, de Kloet ER, Schächinger H & Oitzl MS (2012). Relevance of stress and female sex hormones for emotion and cognition. Cell Mol Neurobiol 32, 725-735.
The Jackson Laboratory (2007). Hematological survey of 11 strains of mice. MPD:Jaxpheno4. The Jackson Laboratory, Mouse Phenome Database web resource.
Tsang JS, Schwartzberg PL, Kotliarov Y, Biancotto A, Xie Z, Germain RN, Wang E, Olnes MJ, Narayanan M, Golding H, Moir S, Dickler HB, Perl S & Cheung F (2014). Global analyses of human immune variation reveal baseline predictors of postvaccination responses. Cell 157, 499-513.
Viselli SM, Reese KR, Fan J, Kovacs WJ & Olsen NJ (1997). Androgens alter B cell development in normal male mice. Cell Immunol 182, 99-104.
Wald C & Wu C (2010). Of Mice and Women: The Bias in Animal Models. Science 327, 1571.
Wallace JG, Potts RH, Szamosi JC, Surette MG & Sloboda DM (2018). The murine female intestinal microbiota does not shift throughout the estrous cycle. PLoS One 13, e0200729-e0200729.
Wilhelmson AS, Lantero Rodriguez M, Stubelius A, Fogelstrand P, Johansson I, Buechler MB, Lianoglou S, Kapoor VN, Johansson ME, Fagman JB, Duhlin A, Tripathi P, Camponeschi A, Porse BT, Rolink AG, Nissbrandt H, Turley SJ, Carlsten H, Martensson IL, Karlsson MCI & Tivesten A (2018). Testosterone is an endogenous regulator of BAFF and splenic B cell number. Nat Commun 9, 2067.
Woitowich NC, Beery A & Woodruff T (2020). A 10-year follow-up study of sex inclusion in the biological sciences. eLife 9, e56344.
Yang H, Bell TA, Churchill GA & Pardo-Manuel de Villena F (2007). On the subspecific origin of the laboratory mouse. Nat Genet 39, 1100-1107.
Yang H, Wang JR, Didion JP, Buus RJ, Bell TA, Welsh CE, Bonhomme F, Yu AH, Nachman MW, Pialek J, Tucker P, Boursot P, McMillan L, Churchill GA & de Villena FP (2011). Subspecific origin and haplotype diversity in the laboratory mouse. Nat Genet 43, 648-655.