Immunohistochemical demonstration of Toll Like Receptors - 2, 4 and 6 in mycoplasmal pneumonia

Gözde Yücel; Arda Selin Tunç; Özgür Özöner; Yavuz Ulusoy; Bahadır Kılınç; Sevil Vural

doi:10.33988/auvfd.1901529

Immunohistochemical demonstration of Toll Like Receptors - 2, 4 and 6 in mycoplasmal pneumonia

Abstract

Mycoplasmal pneumonia is a highly contagious disease that causes serious economic losses in sheep, goats, and cattle worldwide. The aim of this study was to investigate the pathomorphological findings and the immunohistochemical distribution of Toll-Like Receptors (TLRs) involved in the pathogenesis of mycoplasmal pneumonia (n=40). Histopathological examination revealed, purulent bronchopneumonia (n=13), fibrinous bronchopneumonia (n=14), interstitial pneumonia (n=6), granulomatous pneumonia (n=1), and non-specific lesions (n=6). The same tissue sections were immunohistochemically stained with antibodies against Mycoplasma pneumoniae, TLR-2, TLR-4, and TLR-6. Immunopositivity for TLRs was observed in bronchial, bronchiolar, alveolar epithelial cells and inflammatory cells. Although no statistically significant difference was found between the groups, TLR-2-positive cell counts were numerically higher in purulent inflammation, whereas TLR-4-positive cell counts were numerically higher in fibrinous inflammation. The results indicate that TLR-2, TLR-4, and TLR-6 may contribute to the inflammatory response in mycoplasmal pneumonia.

Keywords

Project Number

Ankara University Scientific Research Projects Coordination Unit, Project number: 18H0239001.

References

Akira S, Takeda K (2004): Toll-like receptor signalling. Nat Rev Immunol, 4, 499-511. DOI: 10.1038/nri1391.
Arancibia SA, Beltrán CJ, Aguirre IM, et al (2007): Toll-like receptors are key participants in innate immune responses. Biol Res, 40, 97-112. DOI: 10.4067/s0716-97602007000200001.
Armstrong L, Medford AR, Uppington KM, et al (2004): Expression of functional toll-like receptor-2 and-4 on alveolar epithelial cells. Am J Respir Cell Mol Biol, 31, 241–245. DOI: 10.1165/rcmb.2004-0078oc.
Chen M, Deng, H, Zhao Y, et al (2022): Toll-like receptor 2 modulates pulmonary inflammation and TNF-α release mediated by Mycoplasma pneumoniae. Front Cell Infect Microbiol, 12, 824027. DOI: 10.3389/fcimb.2022.824027.
Cottew GS, Lloyd LC (1965): An outbreak of pleurisy and pneumonia in goats in Australia attributed to a mycoplasma species. J Comp Pathol, 75, 363-374. DOI: 10.1016/0021-9975(65)90016-2.
DaMassa AJ, Wakenell PS, Brooks DL (1992): Mycoplasmas of goats and sheep. J Vet Diagn Invest, 4, 101-113. DOI: 10.1177/104063879200400126
De Nardo D (2015): Toll-like receptors: activation, signalling and transcriptional modulation. Cytokine, 74, 181-189. DOI:10.1016/j.cyto.2015.02.025.
Di Teodoro G, Marruchella G, Di Provvido A, et al (2020): Contagious bovine pleuropneumonia: a comprehensive overview. Vet Pathol, 57, 476-489. DOI: 10.1177/0300985820921818.

Droemann D, Goldmann T, Branscheid D, et al (2003): Toll-like receptor 2 is expressed by alveolar epithelial cells type II and macrophages in the human lung. Histochem Cell Biol, 119, 103-108. DOI: 10.1007/s00418-003-0497-4.
Ferreira DS, Annoni R, Silva LF, et al (2012): Toll–like receptors 2, 3 and 4 and thymic stromal lymphopoietin expression in fatal asthma. Clin Exp Allergy, 42, 1459-1471. DOI: 10.1111/j.1365-2222.2012.04047.x.
Gagea MI, Bateman KG, Shanahan RA, et al (2006): Naturally occurring Mycoplasma bovis—associated pneumonia and polyarthritis in feedlot beef calves. J Vet Diagn Invest, 18, 29-40. DOI: 10.1177/104063870601800105.
Gourlay RN, Thomas LH, Wyld SG (1989): Increased severity of calf pneumonia associated with the appearance of Mycoplasma bovis in a rearing herd. Vet Rec, 124, 420-422. DOI: 10.1136/vr.124.16.420.
Hasso SA, Al-Darraji AM, Al-Aubaidi JM (1994): Pathology of experimentally induced contagious agalactia in goats. Small Rumin Res, 13, 79–84, 1994. DOI: 10.1016/0921-4488(94)90034-5.
Hermeyer K, Buchenau I, Thomasmeyer A, et al (2012): Chronic pneumonia in calves after experimental infection with Mycoplasma bovis strain 1067: characterization of lung pathology, persistence of variable surface protein antigens and local immune response. Acta Vet Scand, 54, 1-11. DOI: 10.1186/1751-0147-54-9.
Hodgkinson CP, Patel K, Ye S (2008): Functional Toll-like receptor 4 mutations modulate the response to fibrinogen. J Thromb Haemost, 100, 301-307. DOI: 10.1160/TH08-03-0179.
Janeway CA, Medzhitov R (2002): Innate immune recognition. Annu Rev Immunol, 20, 197-216. DOI: 10.1146/annurev.immunol.20.083001.084359.
Jiang D, Liang J, Li Y, Noble PW (2006): The role of Toll-like receptors in non-infectious lung injury. Cell Res, 16, 693-701. DOI: 10.1038/sj.cr.7310085.
Waheed Khudiar Khareeb Z, Ismail Ibrahim Z, Ali Abdullah F (2022): Pathological and molecular detection of Mycoplasma ovipneumoniae in sheep, Basrah Province, Iraq. Arch Razi Inst, 77, 2073-2080. DOI: 10.22092/ARI.2022.357996.2134.
Kiechl S, Lorenz E, Reindl M, et al (2002): Toll-like receptor 4 polymorphisms and atherogenesis. N Engl J Med, 347, 185–192. DOI: 10.1056/NEJMoa012673.
Kovach MA, Standiford TJ (2011): Toll like receptors in diseases of the lung. Int Immunopharmacol, 11,1399-1406. DOI: 10.1016/j.intimp.2011.05.013.
Kumar H, Kawai T, Akira S (2009): Toll-like receptors and innate immunity. Biochem Biophys Res Commun, 388, 621–625. DOI: 10.1016/j.bbrc.2009.08.062.
Kumar V, Stewart IV JH (2024): Toll-Like Receptors in Immunity and Inflammation. In Thirty Years since the Discovery of Toll-Like Receptors, IntechOpen. DOI: 10.5772/intechopen.1003992.
Kundakcı A, Pirat A (2012): Toll Benzeri Reseptörler. Turk J Intensive Care, 10, 63-73. DOI: org/10.4274/tybdd. 10.1.
Le Mandat Schultz A, Bonnard A, Barreau F, et al (2007): Expression of TLR-2, TLR-4, NOD2 and pNF-κB in a neonatal rat model of necrotizing enterocolitis. PloS one, 2, e1102. DOI: 10.1371/journal.pone.0001102.
Liu T, Zhang L, Joo D, et al (2017): NF-κB signaling in inflammation. Signal Transduct Target Ther, 2, 1–9. DOI: 10.1038/sigtrans.2017.23.
López A, Martinson SA (2017): Respiratory System, Mediastinum, and Pleurae. 471–540. In: James F. Zachary (Ed), Pathologic Basis of Veterinary Disease – Expert Consult, 6th ed. Saint Louis: Elsevier, Saunders.
Luo H, He J, Qin L, et al (2021): Mycoplasma pneumoniae lipids license TLR-4 for activation of NLRP3 inflammasome and autophagy to evoke a proinflammatory response. Clin Exp Immunol, 203, 66-79. DOI: 10.1111/ cei.13510.
Maris NA, Dessing MC, De Vos AF, et al (2006): Toll-like receptor mRNA levels in alveolar macrophages after inhalation of endotoxin. Eur Respir J, 28, 622-626. DOI: 10.1183/09031936.06.00010806.
Naghib M, Hatam-Jahromi M, Niktab M, et al (2018): Mycoplasma pneumoniae and toll-like receptors: A mutual avenue. Allergol Immunopathol, 46, 508-513. DOI: 10.1016/j.aller.2017.09.021.
Ozinsky A, Underhill DM, Fontenot JD, et al (2000): The repertoire for pattern recognition of pathogens by the innate immune system is defined by cooperation between toll-like receptors. Proc Natl Acad Sci, 97, 13766-13771. DOI: 10.1073/pnas.250476497.
Pelka K, Phulphagar K, Zimmermann J, et al (2014): Cutting edge: the UNC93B1 tyrosine-based motif regulates trafficking and TLR responses via separate mechanisms. J Immunol, 193, 3257-3261. DOI:10.4049/jimmunol. 1301886.
Rodriguez F, Bryson DG, Ball HJ, et al (1996): Pathological and immunohistochemical studies of natural and experimental Mycoplasma bovis pneumonia in calves. J Comp Pathol, 115, 151–162. DOI: 10.1016/S0021-9975(96)80037-5.
Sabroe I, Parker LC, Dower SK, et al (2008): The role of TLR activation in inflammation. J Pathol, 214, 126–135. DOI:10.1002/path.2264.
Saraya T, Nakata K, Nakagaki K, et al (2011): Identification of a mechanism for lung inflammation caused by Mycoplasma pneumoniae using a novel mouse model. Results Immunol, 1, 76-87, 2011. DOI: 10.1016/j.rinim.2011.11.001.
Schmaußer B, Andrulis M, Endrich S, et al (2005): Toll-like receptors TLR4, TLR5 and TLR9 on gastric carcinoma cells: an implication for interaction with Helicobacter pylori. Int J Med Microbiol, 295, 179-185, 2005. DOI: 10.1016/j.ijmm.2005.02.009.
Sha Q, Truong-Tran AQ, Plitt JR, et al (2004): Activation of airway epithelial cells by toll-like receptor agonists. Am J Respir Cell Mol Biol, 31, 358-364. DOI: 10.1165/rcmb.2003-0388OC.
Shin HS, Xu F, Bagchi A, et al (2011): Bacterial lipoprotein TLR2 agonists broadly modulate endothelial function and coagulation pathways in vitro and in vivo. J Immunol, 186, 1119-1130. DOI: 10.4049/jimmunol. 1001647.
Smiley ST, King JA, Hancock WW (2001): Fibrinogen stimulates macrophage chemokine secretion through toll-like receptor 4. J Immunol, 167, 2887-2894. DOI: 10.4049/jimmunol.167.5.2887.
Tabeta K, Hoebe K, Janssen EM, et al (2006): The Unc93b1 mutation 3d disrupts exogenous antigen presentation and signaling via Toll-like receptors 3, 7 and 9. Nat Immunol, 7, 156–164. DOI: 10.1038/ni1297.
Tabeta K, Yamazaki K, Akashi S, et al (2000): Toll-like receptors confer responsiveness to lipopolysaccharide from Porphyromonas gingivalis in human gingival fibroblasts. Infect Immun, 68, 3731-3735. DOI: 10.1128/iai.68.6.3731-3735.
Taguchi T, Mukai K (2019): Innate immunity signalling and membrane trafficking. Curr Opin Cell Biol, 59, 1–7. DOI: 10.1016/j.ceb.2019.02.002.
Takeuchi O, Kaufmann A, Grote K, et al (2000): Cutting edge: preferentially the R-stereoisomer of the mycoplasmal lipopeptide macrophage-activating lipopeptide-2 activates immune cells through a toll-like receptor 2-and MyD88-dependent signaling pathway. J Immunol, 164, 554–557. DOI: 10.4049/jimmunol.164.2.554. v
Thomas LH, Howard CJ, Stott EJ, et al (1986): Mycoplasma bovis infection in gnotobiotic calves and combined infection with respiratory syncytial virus. Vet Pathol, 23, 571–578. DOI:10.1177/030098588602300505.
Thoma-Uszynski S, Stenger S, Takeuchi O, et al (2001): Induction of direct antimicrobial activity through mammalian toll-like receptors. Science, 291, 1544-1547. DOI: 10.1126/science.291.5508.1544.
Underhill DM, Ozinsky A (2002): Phagocytosis of microbes: complexity in action. Annu Rev Immunol, 20, 825–852. DOI: 10.1146/annurev.immunol.20.103001. 114744.
Wang PL, Azuma Y, Shinohara M, et al (2000): Toll-like receptor 4-mediated signal pathway induced by Porphyromonas gingivalis lipopolysaccharide in human gingival fibroblasts. Biochem Biophys Res Commun, 273, 1161-1167. DOI: 10.1006/bbrc.2000.3060.
Xue D, Ma Y, Li M, et al (2015): Mycoplasma ovipneumoniae induces inflammatory response in sheep airway epithelial cells via a MyD88-dependent TLR signaling pathway. Vet Immunol Immunopathol. 163, 57-66. DOI: 10.1016/j.vetimm.2014.11.008.

Details

Primary Language

English

Subjects

Veterinary Pathology

Journal Section

Research Article

Authors

Gözde Yücel ^*
0000-0002-2586-8346
Türkiye

Arda Selin Tunç
0000-0002-4813-7626
Türkiye

Özgür Özöner
0000-0001-7354-0655
Türkiye

Yavuz Ulusoy This is me
0000-0001-6942-5013
Türkiye

Bahadır Kılınç
0000-0000-0000-0000
Türkiye

Sevil Vural
0000-0003-2111-3381
Türkiye

Early Pub Date

September 18, 2026

Publication Date

-

Submission Date

March 14, 2026

Acceptance Date

July 3, 2026

Published in Issue

Year 2026 Number: Advanced Online Publication

DOI

https://doi.org/10.33988/auvfd.1901529

IZ

https://izlik.org/JA84XC25SD

Cite

RIS / Bibtex

APA

Yücel, G., Tunç, A. S., Özöner, Ö., Ulusoy, Y., Kılınç, B., & Vural, S. (2026). Immunohistochemical demonstration of Toll Like Receptors - 2, 4 and 6 in mycoplasmal pneumonia. Ankara Üniversitesi Veteriner Fakültesi Dergisi, Advanced Online Publication. https://doi.org/10.33988/auvfd.1901529

AMA

1.Yücel G, Tunç AS, Özöner Ö, Ulusoy Y, Kılınç B, Vural S. Immunohistochemical demonstration of Toll Like Receptors - 2, 4 and 6 in mycoplasmal pneumonia. Ankara Univ Vet Fak Derg. 2026;(Advanced Online Publication). doi:10.33988/auvfd.1901529

Chicago

Yücel, Gözde, Arda Selin Tunç, Özgür Özöner, Yavuz Ulusoy, Bahadır Kılınç, and Sevil Vural. 2026. “Immunohistochemical Demonstration of Toll Like Receptors - 2, 4 and 6 in Mycoplasmal Pneumonia”. Ankara Üniversitesi Veteriner Fakültesi Dergisi, no. Advanced Online Publication. https://doi.org/10.33988/auvfd.1901529.

EndNote

Yücel G, Tunç AS, Özöner Ö, Ulusoy Y, Kılınç B, Vural S (September 1, 2026) Immunohistochemical demonstration of Toll Like Receptors - 2, 4 and 6 in mycoplasmal pneumonia. Ankara Üniversitesi Veteriner Fakültesi Dergisi Advanced Online Publication

IEEE

[1]G. Yücel, A. S. Tunç, Ö. Özöner, Y. Ulusoy, B. Kılınç, and S. Vural, “Immunohistochemical demonstration of Toll Like Receptors - 2, 4 and 6 in mycoplasmal pneumonia”, Ankara Univ Vet Fak Derg, no. Advanced Online Publication, Sept. 2026, doi: 10.33988/auvfd.1901529.

ISNAD

Yücel, Gözde - Tunç, Arda Selin - Özöner, Özgür - Ulusoy, Yavuz - Kılınç, Bahadır - Vural, Sevil. “Immunohistochemical Demonstration of Toll Like Receptors - 2, 4 and 6 in Mycoplasmal Pneumonia”. Ankara Üniversitesi Veteriner Fakültesi Dergisi. Advanced Online Publication (September 1, 2026). https://doi.org/10.33988/auvfd.1901529.

JAMA

1.Yücel G, Tunç AS, Özöner Ö, Ulusoy Y, Kılınç B, Vural S. Immunohistochemical demonstration of Toll Like Receptors - 2, 4 and 6 in mycoplasmal pneumonia. Ankara Univ Vet Fak Derg. 2026. doi:10.33988/auvfd.1901529.

MLA

Yücel, Gözde, et al. “Immunohistochemical Demonstration of Toll Like Receptors - 2, 4 and 6 in Mycoplasmal Pneumonia”. Ankara Üniversitesi Veteriner Fakültesi Dergisi, no. Advanced Online Publication, Sept. 2026, doi:10.33988/auvfd.1901529.

Vancouver

1.Gözde Yücel, Arda Selin Tunç, Özgür Özöner, Yavuz Ulusoy, Bahadır Kılınç, Sevil Vural. Immunohistochemical demonstration of Toll Like Receptors - 2, 4 and 6 in mycoplasmal pneumonia. Ankara Univ Vet Fak Derg. 2026 Sep. 1;(Advanced Online Publication). doi:10.33988/auvfd.1901529