Repeated Inhalation of GM-CSF by Nonhuman Primates Induces Bronchus-associated Lymphoid Tissue Along the Lower Respiratory Tract

Overview

Journal Respir Res

Specialty Pulmonary Medicine

Date 2024 Nov 10

PMID 39523334

Authors

Ryushi Tazawa

Riuko Ohashi

Nobutaka Kitamura

Takahiro Tanaka

Kazuhide Nakagaki

Sachiko Yuki

Atsushi Fujiwara

Koh Nakata

Affiliations

Soon will be listed here.

Abstract

Background: Repeated inhalation of granulocyte-macrophage colony-stimulating factor (GM-CSF) was recently approved in Japan as a treatment for autoimmune pulmonary alveolar proteinosis. However, the detailed physiological and pathological effects of repeated inhalation in the long term, especially at increasing doses, remain unclear.

Methods: In this chronic safety study, we administered 24 cynomolgus monkeys (Macaca fascicularis) aged 2-3 years with aerosolized sargramostim (a yeast-derived recombinant human GM-CSF [rhGM-CSF]) biweekly for 26 weeks across four dosing groups (0, 5, 100, and 500 µg/kg/day). We measured the serum GM-CSF antibody (GM-Ab) concentration by an ELISA and assessed the neutralizing capacity of GM-Ab using the GM-CSF-dependent cell line TF-1. We subjected lung tissue samples taken from all monkeys at 27 weeks to histopathological assessment using a sargramostim-specific monoclonal antibody to detect localization of residual sargramostim.

Results: All the animals maintained good body condition and showed steady weight gain throughout the study. The pathological analyses of the lung revealed the formation of induced bronchus-associated lymphoid tissue (iBALT) in the lower respiratory tract, even at the clinical dose of 5 µg/kg/day. There was a relationship between the number or size of BALT and sargramostim dose or the serum GM-Ab levels. Immunohistochemical analyses revealed GM-Ab-producing cells in the follicular region of iBALT, with residual sargramostim in the follicles. Leucocyte counts were inversely correlated with GM-Ab levels in the high-dose groups. Additionally, serum GM-Ab from the treated animals significantly suppressed the alveolar macrophage proliferation activity of both Cynomolgus recombinant and rhGM-CSF in vitro.

Conclusion: Long-term repeated inhalation of sargramostim led to iBALT formation in the lower respiratory tract, even at the clinical dose of 5 µg/kg/day, with the extent of iBALT formation increasing in a dose-dependent manner. Inhaled sargramostim was localized to the follicular region of iBALT nodules, which may induce the production of GM-Ab.

References

Nei T, Urano S, Motoi N, Hashimoto A, Kitamura N, Tanaka T . Memory B cell pool of autoimmune pulmonary alveolar proteinosis patients contains higher frequency of GM-CSF autoreactive B cells than healthy subjects. Immunol Lett. 2019; 212:22-29. DOI: 10.1016/j.imlet.2019.05.013. View

Tazawa R, Ueda T, Abe M, Tatsumi K, Eda R, Kondoh S . Inhaled GM-CSF for Pulmonary Alveolar Proteinosis. N Engl J Med. 2019; 381(10):923-932. DOI: 10.1056/NEJMoa1816216. View

Pabst R, Gehrke I . Is the bronchus-associated lymphoid tissue (BALT) an integral structure of the lung in normal mammals, including humans?. Am J Respir Cell Mol Biol. 1990; 3(2):131-5. DOI: 10.1165/ajrcmb/3.2.131. View

Thomson R, Loebinger M, Burke A, Morgan L, Waterer G, Ganslandt C . OPTIMA: An Open-Label, Noncomparative Pilot Trial of Inhaled Molgramostim in Pulmonary Nontuberculous Mycobacterial Infection. Ann Am Thorac Soc. 2023; 21(4):568-576. DOI: 10.1513/AnnalsATS.202306-532OC. View

Bosteels C, Van Damme K, De Leeuw E, Declercq J, Maes B, Bosteels V . Loss of GM-CSF-dependent instruction of alveolar macrophages in COVID-19 provides a rationale for inhaled GM-CSF treatment. Cell Rep Med. 2022; 3(12):100833. PMC: 9663750. DOI: 10.1016/j.xcrm.2022.100833. View

Zsengeller Z, Reed J, Bachurski C, Levine A, Hirsch R, Whitsett J . Adenovirus-mediated granulocyte-macrophage colony-stimulating factor improves lung pathology of pulmonary alveolar proteinosis in granulocyte-macrophage colony-stimulating factor-deficient mice. Hum Gene Ther. 1998; 9(14):2101-9. DOI: 10.1089/hum.1998.9.14-2101. View

Shima K, Arumugam P, Sallese A, Horio Y, Ma Y, Trapnell C . A murine model of hereditary pulmonary alveolar proteinosis caused by homozygous gene disruption. Am J Physiol Lung Cell Mol Physiol. 2022; 322(3):L438-L448. PMC: 8917935. DOI: 10.1152/ajplung.00175.2021. View

Suzuki T, Trapnell B . Pulmonary Alveolar Proteinosis Syndrome. Clin Chest Med. 2016; 37(3):431-40. PMC: 5902187. DOI: 10.1016/j.ccm.2016.04.006. View

Suda T, Chida K, Hayakawa H, Imokawa S, Iwata M, Nakamura H . Development of bronchus-associated lymphoid tissue in chronic hypersensitivity pneumonitis. Chest. 1999; 115(2):357-63. DOI: 10.1378/chest.115.2.357. View

10.

Paine R, Chasse R, Halstead E, Nfonoyim J, Park D, Byun T . Inhaled Sargramostim (Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor) for COVID-19-Associated Acute Hypoxemia: Results of the Phase 2, Randomized, Open-Label Trial (iLeukPulm). Mil Med. 2022; 188(7-8):e2629-e2638. PMC: 10363010. DOI: 10.1093/milmed/usac362. View

11.

Sato A, Chida K, Iwata M, Hayakawa H . Study of bronchus-associated lymphoid tissue in patients with diffuse panbronchiolitis. Am Rev Respir Dis. 1992; 146(2):473-8. DOI: 10.1164/ajrccm/146.2.473. View

12.

Tanaka N, Newell J, Brown K, Cool C, Lynch D . Collagen vascular disease-related lung disease: high-resolution computed tomography findings based on the pathologic classification. J Comput Assist Tomogr. 2004; 28(3):351-60. DOI: 10.1097/00004728-200405000-00009. View

13.

Naito M . Macrophage differentiation and function in health and disease. Pathol Int. 2008; 58(3):143-55. DOI: 10.1111/j.1440-1827.2007.02203.x. View

14.

Raghu G, Remy-Jardin M, Ryerson C, Myers J, Kreuter M, Vasakova M . Diagnosis of Hypersensitivity Pneumonitis in Adults. An Official ATS/JRS/ALAT Clinical Practice Guideline. Am J Respir Crit Care Med. 2020; 202(3):e36-e69. PMC: 7397797. DOI: 10.1164/rccm.202005-2032ST. View

15.

Sato A, Hayakawa H, Uchiyama H, Chida K . Cellular distribution of bronchus-associated lymphoid tissue in rheumatoid arthritis. Am J Respir Crit Care Med. 1996; 154(6 Pt 1):1903-7. DOI: 10.1164/ajrccm.154.6.8970384. View

16.

Garg D, Mody M, Pal C, Patel P, Migliore C, Minerowicz C . Follicular Bronchiolitis: Two Cases with Varying Clinical and Radiological Presentation. Case Rep Pulmonol. 2020; 2020:4564587. PMC: 7007759. DOI: 10.1155/2020/4564587. View

17.

Markovic S, Suman V, Nevala W, Geeraerts L, Creagan E, Erickson L . A dose-escalation study of aerosolized sargramostim in the treatment of metastatic melanoma: an NCCTG Study. Am J Clin Oncol. 2008; 31(6):573-9. PMC: 2694721. DOI: 10.1097/COC.0b013e318173a536. View

18.

Weinman J, Manning D, Liptzin D, Krausert A, Browne L . HRCT findings of childhood follicular bronchiolitis. Pediatr Radiol. 2017; 47(13):1759-1765. DOI: 10.1007/s00247-017-3951-5. View

19.

Ragnhammar P, Friesen H, Frodin J, Lefvert A, Hassan M, Osterborg A . Induction of anti-recombinant human granulocyte-macrophage colony-stimulating factor (Escherichia coli-derived) antibodies and clinical effects in nonimmunocompromised patients. Blood. 1994; 84(12):4078-87. View

20.

Hayakawa H, Sato A, Imokawa S, Toyoshima M, Chida K, Iwata M . Bronchiolar disease in rheumatoid arthritis. Am J Respir Crit Care Med. 1996; 154(5):1531-6. DOI: 10.1164/ajrccm.154.5.8912776. View