Background: Hofbauer cells (HBCs) are fetal-origin macrophages in the placental villous stroma that contribute to immune tolerance at the feto-maternal interface. They predominantly display an M2 phenotype, characterized by CD206 expression. Methods: Using immunohistochemistry and morphometric analysis, we quantified HBCs, assessed CD206 intensity and morphology, and evaluated apoptotic body accumulation in placental villi. Comparisons were made among pregnancies complicated by type 1 diabetes mellitus (T1DM), gestational diabetes mellitus (GDM), and normoglycemic controls, as well as between male and female fetuses. Results: Significant effects of maternal diabetes and fetal sex on CD206 intensity were observed ([diagnosis: F = 2773.00, p < 0.0001; sex: F = 12.19, p = 0.0005]), with a strong interaction (F = 165.40, p < 0.0001). In controls, CD206 intensity was higher in female than male fetuses (p < 0.0001). Across groups, CD206 intensity decreased progressively from controls to GDM and T1DM, with a more pronounced reduction in females. Reduced CD206 was associated with elongation and irregular HBC morphology and increased IL-1β (r = -0.392, p = 0.003; r = -0.609, p < 0.0001) suggesting less tolerogenic phenotype. For apoptotic bodies, significant main effects of maternal diabetes and fetal sex were detected ([diagnosis: F = 97.16, p < 0.0001; sex: F = 15.88, p = 0.0001]). Accumulation increased progressively from controls to GDM and T1DM, with higher counts in males. Conclusions: Maternal diabetes is associated with reduced CD206 intensity, altered HBC morphology, and accumulation of apoptotic bodies in placental villi. Our results suggest greater HBC plasticity, potentially contributing to a tolerogenic placental environment in females.

Department of Histology and Embryology Faculty of Medicine and Dentistry Palacky University 779 00 Olomouc Czech Republic

Zobrazit více v PubMed

Ingman K., Cookson V.J., Jones C.J., Aplin J.D. Characterisation of Hofbauer cells in first and second trimester placenta: Incidence, phenotype, survival in vitro and motility. Placenta. 2010;31:535–544. doi: 10.1016/j.placenta.2010.03.003. PubMed DOI

Loegl J., Hiden U., Nussbaumer E., Schliefsteiner C., Cvitic S., Lang I., Wadsack C., Huppertz B., Desoye G. Hofbauer cells of M2a, M2b and M2c polarization may regulate feto-placental angiogenesis. Reproduction. 2016;152:447–455. doi: 10.1530/REP-16-0159. PubMed DOI

Tang Z., Buhimschi I.A., Buhimschi C.S., Tadesse S., Norwitz E., Niven-Fairchild T., Huang S.T., Guller S. Decreased levels of folate receptor-β and reduced numbers of fetal macrophages (Hofbauer cells) in placentas from pregnancies with severe pre-eclampsia. Am. J. Reprod. Immunol. 2013;70:104–115. doi: 10.1111/aji.12112. PubMed DOI PMC

Schliefsteiner C., Peinhaupt M., Kopp S., Lögl J., Lang-Olip I., Hiden U., Heinemann A., Desoye G., Wadsack C. Human Placental Hofbauer Cells Maintain an Anti-inflammatory M2 Phenotype despite the Presence of Gestational Diabetes Mellitus. Front. Immunol. 2017;8:888. doi: 10.3389/fimmu.2017.00888. PubMed DOI PMC

Swieboda D., Johnson E.L., Beaver J., Haddad L., Enninga E.A.L., Hathcock M., Cordes S., Jean V., Lane I., Skountzou I., et al. Baby’s First Macrophage: Temporal Regulation of Hofbauer Cell Phenotype Influences Ligand-Mediated Innate Immune Responses across Gestation. J. Immunol. 2020;204:2380–2391. doi: 10.4049/jimmunol.1901185. PubMed DOI PMC

Thomas J.R., Appios A., Zhao X., Dutkiewicz R., Donde M., Lee C.Y.C., Naidu P., Lee C., Cerveira J., Liu B., et al. Phenotypic and functional characterization of first-trimester human placental macrophages, Hofbauer cells. J. Exp. Med. 2021;218:e20200891. doi: 10.1084/jem.20200891. PubMed DOI PMC

Yao Y., Xu X.H., Jin L. Macrophage Polarization in Physiological and Pathological Pregnancy. Front. Immunol. 2019;10:792. doi: 10.3389/fimmu.2019.00792. PubMed DOI PMC

Nielsen M.C., Andersen M.N., Grønbæk H., Damgaard Sandahl T., Møller H.J. Extracellular vesicle-associated soluble CD163 and CD206 in patients with acute and chronic inflammatory liver disease. Scand. J. Gastroenterol. 2020;55:588–596. doi: 10.1080/00365521.2020.1759140. PubMed DOI

Taylor P.R., Gordon S., Martinez-Pomares L. The mannose receptor: Linking homeostasis and immunity through sugar recognition. Trends Immunol. 2005;26:104–110. doi: 10.1016/j.it.2004.12.001. PubMed DOI

Sturge J., Todd S.K., Kogianni G., McCarthy A., Isacke C.M. Mannose receptor regulation of macrophage cell migration. J. Leukoc. Biol. 2007;82:585–593. doi: 10.1189/jlb.0107053. PubMed DOI

Jones G.E. Cellular signaling in macrophage migration and chemotaxis. J. Leukoc. Biol. 2000;68:593–602. doi: 10.1189/jlb.68.5.593. PubMed DOI

Qaseem M., Ara N., Farooq K., Idrees S.A. Histological Changes in the Extracellular Matrix of the Human Placenta Complicated by Diabetes. Med. Forum Mon. 2024;35:57–61.

Hod M., Kapur A., Sacks D.A., Hadar E., Agarwal M., Di Renzo G.C., Cabero Roura L., McIntyre H.D., Morris J.L., Divakar H. The International Federation of Gynecology and Obstetrics (FIGO) Initiative on gestational diabetes mellitus: A pragmatic guide for diagnosis, management, and care. Int. J. Gynaecol. Obstet. 2015;131((Suppl. 3)):S173–S211. doi: 10.1016/S0020-7292(15)30007-2. PubMed DOI

Kilpeläinen T.O., Zillikens M.C., Stančákova A., Finucane F.M., Ried J.S., Langenberg C., Zhang W., Beckmann J.S., Luan J., Vandenput L., et al. Genetic variation near IRS1 associates with reduced adiposity and an impaired metabolic profile. Nat. Genet. 2011;43:753–760. doi: 10.1038/ng.866. PubMed DOI PMC

Radaelli T., Varastehpour A., Catalano P., Hauguel-de Mouzon S. Gestational diabetes induces placental genes for chronic stress and inflammatory pathways. Diabetes. 2003;52:2951–2958. doi: 10.2337/diabetes.52.12.2951. PubMed DOI

Cano-Cano F., Gómez-Jaramillo L., Ramos-García P., Arroba A.I., Aguilar-Diosdado M. IL-1β Implications in Type 1 Diabetes Mellitus Progression: Systematic Review and Meta-Analysis. J. Clin. Med. 2022;11:1303. doi: 10.3390/jcm11051303. PubMed DOI PMC

Wen Y., Gu J., Li S.-L., Reddy M.A., Natarajan R., Nadler J.L. Elevated Glucose and Diabetes Promote Interleukin-12 Cytokine Gene Expression in Mouse Macrophages. Endocrinology. 2006;147:2518–2525. doi: 10.1210/en.2005-0519. PubMed DOI

Sisino G., Bouckenooghe T., Aurientis S., Fontaine P., Storme L., Vambergue A. Diabetes during pregnancy influences Hofbauer cells, a subtype of placental macrophages, to acquire a pro-inflammatory phenotype. Biochim. Biophys. Acta. 2013;1832:1959–1968. doi: 10.1016/j.bbadis.2013.07.009. PubMed DOI

Dai M., Wu L., Wang P., Wen Z., Xu X., Wang D.W. CYP2J2 and Its Metabolites EETs Attenuate Insulin Resistance via Regulating Macrophage Polarization in Adipose Tissue. Sci. Rep. 2017;7:46743. doi: 10.1038/srep46743. PubMed DOI PMC

Orliaguet L., Ejlalmanesh T., Alzaid F. Metabolic and Molecular Mechanisms of Macrophage Polarisation and Adipose Tissue Insulin Resistance. Int. J. Mol. Sci. 2020;21:5731. doi: 10.3390/ijms21165731. PubMed DOI PMC

Tauber Z., Burianova A., Koubova K., Mrstik M., Jirkovska M., Cizkova K. The interplay of inflammation and placenta in maternal diabetes: Insights into Hofbauer cell expression patterns. Front. Immunol. 2024;15:1386528. doi: 10.3389/fimmu.2024.1386528. PubMed DOI PMC

Baines K.J., West R.C. Sex differences in innate and adaptive immunity impact fetal, placental, and maternal health. Biol. Reprod. 2023;109:256–270. doi: 10.1093/biolre/ioad072. PubMed DOI

Pantazi P., Kaforou M., Tang Z., Abrahams V.M., McArdle A., Guller S., Holder B. Placental macrophage responses to viral and bacterial ligands and the influence of fetal sex. iScience. 2022;25:105653. doi: 10.1016/j.isci.2022.105653. PubMed DOI PMC

Batorsky R., Ceasrine A.M., Shook L.L., Kislal S., Bordt E.A., Devlin B.A., Perlis R.H., Slonim D.K., Bilbo S.D., Edlow A.G. Hofbauer cells and fetal brain microglia share transcriptional profiles and responses to maternal diet-induced obesity. Cell Rep. 2024;43:114326. doi: 10.1016/j.celrep.2024.114326. PubMed DOI PMC

Landini G., Martinelli G., Piccinini F. Colour deconvolution: Stain unmixing in histological imaging. Bioinformatics. 2020;37:1485–1487. doi: 10.1093/bioinformatics/btaa847. PubMed DOI

Nguyen D. Quantifying Chromogen Intensity in Immunohistochemistry via Reciprocal Intensity. 2013. [(accessed on 9 September 2024)]. Available online: https://www.protocols.io/view/quantifying-chromogen-intensity-in-immunohistochem-261gerbp7l47/v1/references.

Zhang M., Cui D., Yang H. The Distributional Characteristics of M2 Macrophages in the Placental Chorionic Villi are Altered Among the Term Pregnant Women with Uncontrolled Type 2 Diabetes Mellitus. Front. Immunol. 2022;13:837391. doi: 10.3389/fimmu.2022.837391. PubMed DOI PMC

Gosain R., Motwani R., Anupama H. CD68 expression in the placenta of gestational diabetic mothers: A case-control study. Indian J. Pathol. Microbiol. 2023;66:727–731. doi: 10.4103/ijpm.ijpm_99_22. PubMed DOI

Kerby A., Shingleton D., Batra G., Sharps M.C., Baker B.C., Heazell A.E.P. Placental Morphology and Cellular Characteristics in Stillbirths in Women with Diabetes and Unexplained Stillbirths. Arch. Pathol. Lab. Med. 2020;145:82–89. doi: 10.5858/arpa.2019-0524-OA. PubMed DOI

Bari M.F., Weickert M.O., Sivakumar K., James S.G., Snead D.R., Tan B.K., Randeva H.S., Bastie C.C., Vatish M. Elevated soluble CD163 in gestational diabetes mellitus: Secretion from human placenta and adipose tissue. PLoS ONE. 2014;9:e101327. doi: 10.1371/journal.pone.0101327. PubMed DOI PMC

Aşır F., Nergiz Y., Nergiz Öztürk Ş., Şahin A., Ağaçayak E. Investigation of placental Hofbauer cells by immunohistochemistry method in complicated pregnancies. Middle East J. Sci. 2021;7:150–159. doi: 10.51477/mejs.1004971. DOI

Dairi A.S., Himayda A.S.A., Moulana A.A.R., Bukhari H.S.H., Hakeem I.M., Elbarrany W. The Effect of Gestational Diabetes Mellitus on the Chorionic Villi of Human Placenta Among Saudi Arabian Mothers: A Quantitative and Comparative Study. Cureus. 2020;12:e11130. doi: 10.7759/cureus.11130. PubMed DOI PMC

Ning J., Zhang M., Cui D., Yang H. The pathologic changes of human placental macrophages in women with hyperglycemia in pregnancy. Placenta. 2022;130:60–66. doi: 10.1016/j.placenta.2022.11.004. PubMed DOI

Paparini D.E., Grasso E., Aguilera F., Arslanian M.A., Lella V., Lara B., Schafir A., Gori S., Merech F., Hauk V., et al. Sex-specific phenotypical, functional and metabolic profiles of human term placenta macrophages. Biol. Sex Differ. 2024;15:80. doi: 10.1186/s13293-024-00652-w. PubMed DOI PMC

McWhorter F.Y., Wang T., Nguyen P., Chung T., Liu W.F. Modulation of macrophage phenotype by cell shape. Proc. Natl. Acad. Sci. USA. 2013;110:17253–17258. doi: 10.1073/pnas.1308887110. PubMed DOI PMC

Eligini S., Crisci M., Bono E., Songia P., Tremoli E., Colombo G.I., Colli S. Human monocyte-derived macrophages spontaneously differentiated in vitro show distinct phenotypes. J. Cell. Physiol. 2013;228:1464–1472. doi: 10.1002/jcp.24301. PubMed DOI

Vogel D.Y., Glim J.E., Stavenuiter A.W., Breur M., Heijnen P., Amor S., Dijkstra C.D., Beelen R.H. Human macrophage polarization in vitro: Maturation and activation methods compared. Immunobiology. 2014;219:695–703. doi: 10.1016/j.imbio.2014.05.002. PubMed DOI

Tedesco S., Bolego C., Toniolo A., Nassi A., Fadini G.P., Locati M., Cignarella A. Phenotypic activation and pharmacological outcomes of spontaneously differentiated human monocyte-derived macrophages. Immunobiology. 2015;220:545–554. doi: 10.1016/j.imbio.2014.12.008. PubMed DOI

Rőszer T. Understanding the Mysterious M2 Macrophage through Activation Markers and Effector Mechanisms. Mediat. Inflamm. 2015;2015:816460. doi: 10.1155/2015/816460. PubMed DOI PMC

van der Zande H.J.P., Nitsche D., Schlautmann L., Guigas B., Burgdorf S. The Mannose Receptor: From Endocytic Receptor and Biomarker to Regulator of (Meta)Inflammation. Front. Immunol. 2021;12:765034. doi: 10.3389/fimmu.2021.765034. PubMed DOI PMC

Jaynes J.M., Sable R., Ronzetti M., Bautista W., Knotts Z., Abisoye-Ogunniyan A., Li D., Calvo R., Dashnyam M., Singh A., et al. Mannose receptor (CD206) activation in tumor-associated macrophages enhances adaptive and innate antitumor immune responses. Sci. Transl. Med. 2020;12:eaax6337. doi: 10.1126/scitranslmed.aax6337. PubMed DOI PMC

Franco-Barraza J., Valdivia-Silva J.E., Zamudio-Meza H., Castillo A., García-Zepeda E.A., Benítez-Bribiesca L., Meza I. Actin cytoskeleton participation in the onset of IL-1beta induction of an invasive mesenchymal-like phenotype in epithelial MCF-7 cells. Arch. Med. Res. 2010;41:170–181. doi: 10.1016/j.arcmed.2010.04.010. PubMed DOI

Kesapragada M., Sun Y.H., Zhu K., Recendez C., Fregoso D., Yang H.Y., Rolandi M., Isseroff R., Zhao M., Gomez M. A data-driven approach to establishing cell motility patterns as predictors of macrophage subtypes and their relation to cell morphology. PLoS ONE. 2024;19:e0315023. doi: 10.1371/journal.pone.0315023. PubMed DOI PMC

Stinson M.W., Liu S., Laurenson A.J., Rotty J.D. Macrophage migration is differentially regulated by fibronectin and laminin through altered adhesion and myosin II localization. Mol. Biol. Cell. 2024;35:ar22. doi: 10.1091/mbc.E23-04-0137. PubMed DOI PMC

Sheng Y.R., Hu W.T., Wei C.Y., Tang L.L., Liu Y.K., Liu Y.Y., Qiu J.P., Li D.J., Zhu X.Y. Insights of efferocytosis in normal and pathological pregnancy. Am. J. Reprod. Immunol. 2019;82:e13088. doi: 10.1111/aji.13088. PubMed DOI

Tao H., Ma R., Cui J., Yang Z., He W., Li Y., Zhao Y. Immunomodulatory effect of efferocytosis at the maternal-fetal interface. Cell Commun. Signal. 2025;23:49. doi: 10.1186/s12964-025-02055-9. PubMed DOI PMC

Yu J., Su X.-L., Jia J., Zeng Y., Zhang J.-Y., Wang S.-S., Feng L., Pan Y., Shi D.-D. The Relationship Between the Expression of Resistin and Apoptosis Factors in Placenta and the Pathogenesis of Gestational Diabetes Mellitus. Matern. Fetal Med. 2020;2:80–83. doi: 10.1097/FM9.0000000000000040. DOI

Sgarbosa F., Barbisan L.F., Brasil M.A., Costa E., Calderon I.M., Gonçalves C.R., Bevilacqua E., Rudge M.V. Changes in apoptosis and Bcl-2 expression in human hyperglycemic, term placental trophoblast. Diabetes Res. Clin. Pract. 2006;73:143–149. doi: 10.1016/j.diabres.2005.12.014. PubMed DOI

Magee T.R., Ross M.G., Wedekind L., Desai M., Kjos S., Belkacemi L. Gestational diabetes mellitus alters apoptotic and inflammatory gene expression of trophobasts from human term placenta. J. Diabetes Complicat. 2014;28:448–459. doi: 10.1016/j.jdiacomp.2014.03.010. PubMed DOI PMC

Hung T.-H., Huang S.-Y., Chen S.-F., Wu C.-P., Hsieh T.S.-T.A. Decreased placental apoptosis and autophagy in pregnancies complicated by gestational diabetes with large-for-gestational age fetuses. Placenta. 2020;90:27–36. doi: 10.1016/j.placenta.2019.12.003. PubMed DOI

Chakraborty B., Chakraborty P., Brown M.C., Crowder D., Goyal A., Safi R., Artham S., Kirkland M., Racioppi A., Juras P., et al. Estrogens increase cancer cell efferocytosis to establish an immunosuppressive tumor microenvironment. bioRxiv. 2024 doi: 10.1101/2024.12.26.630419. PubMed DOI PMC

Xiao Y., Chen Y., Huang S., He H., Hu N., Lin S., You Z. The reduction of microglial efferocytosis is concomitant with depressive-like behavior in CUMS-treated mice. J. Affect. Disord. 2024;352:76–86. doi: 10.1016/j.jad.2024.02.045. PubMed DOI

Jabeen S., Landazuri J., Nagvenkar S., Czuj B., Maghsoudi A., Javdan M., Entezari M., Lockshin R.A., Zakeri Z. TLR4 sex dimorphism correlates with sex dimorphic phagocytosis in primary macrophages. Ital. J. Gend. Specif. Med. 2020;6:100–106. doi: 10.1723/3432.34214. DOI

Igarashi Y., Nawaz A., Kado T., Bilal M., Kuwano T., Yamamoto S., Sasahara M., Jiuxiang X., Inujima A., Koizumi K., et al. Partial depletion of CD206-positive M2-like macrophages induces proliferation of beige progenitors and enhances browning after cold stimulation. Sci. Rep. 2018;8:14567. doi: 10.1038/s41598-018-32803-6. PubMed DOI PMC

Ray A., Hu K.H., Kersten K., Courau T., Kuhn N.F., Zaleta-Linares I., Samad B., Combes A.J., Krummel M.F. Targeting CD206+ macrophages disrupts the establishment of a key antitumor immune axis. J. Exp. Med. 2025;222:e20240957. doi: 10.1084/jem.20240957. PubMed DOI PMC

Akram K.M., Frost L.I., Anumba D.O. Impaired autophagy with augmented apoptosis in a Th1/Th2-imbalanced placental micromilieu is associated with spontaneous preterm birth. Front. Mol. Biosci. 2022;9:897228. doi: 10.3389/fmolb.2022.897228. PubMed DOI PMC

Peelen M.J., Kazemier B.M., Ravelli A.C., De Groot C.J., Van Der Post J.A., Mol B.W., Hajenius P.J., Kok M. Impact of fetal gender on the risk of preterm birth, a national cohort study. Acta Obstet. Gynecol. Scand. 2016;95:1034–1041. doi: 10.1111/aogs.12929. PubMed DOI

Zhou J., Teng Y., Zhang F., Ru X., Li P., Wang J., Yan S., Zhu P., Tao F., Huang K. Sex-specific association between placental inflammatory cytokine mRNA expression and preschoolers’ behavioral development: The Ma’anshan birth cohort study. Brain Behav. Immun. 2022;104:110–121. doi: 10.1016/j.bbi.2022.05.017. PubMed DOI

Fitzgerald E., Shen M., Yong H.E.J., Wang Z., Pokhvisneva I., Patel S., O’Toole N., Chan S.-Y., Chong Y.S., Chen H., et al. Hofbauer cell function in the term placenta associates with adult cardiovascular and depressive outcomes. Nat. Commun. 2023;14:7120. doi: 10.1038/s41467-023-42300-8. PubMed DOI PMC

Tauber Z., Stetkova I., Cizkova K. Influence of fixation method and duration of archiving on immunohistochemical staining intensity in embryonic and fetal tissues. Ann. Anat. Anat. Anz. 2019;224:55–61. doi: 10.1016/j.aanat.2019.03.013. PubMed DOI