Characterization of cytokine profiles and double-positive lymphocyte subpopulations in normal bovine lungs

Nathalie L. Mathy From the Centre for Animal Biotechnology, the University of Melbourne, Parkville, Victoria 3052, Australia.

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John Walker From the Centre for Animal Biotechnology, the University of Melbourne, Parkville, Victoria 3052, Australia.

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Rogan P. Lee From the Centre for Animal Biotechnology, the University of Melbourne, Parkville, Victoria 3052, Australia.

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Abstract

Objective

To characterize cytokine profiles and lymphocyte subpopulations in lung parenchyma and bronchoalveolar lavage (BAL) fluid from normal bovine lungs.

Animals

Eight 12- to 18-month-old cattle.

Procedure

Cell populations in BAL fluid and collagenase-digested lung parenchyma were analyzed by flow cytometry and monoclonal antibodies. Proportions of total cell populations were determined, using Giemsa-stained cytospots. Distribution of lymphocytes within the lung parenchyma was analyzed by immunohistochemistry, and cytokine mRNA species in the parenchyma were characterized by use of reverse transcriptase-polymerase chain reaction analysis.

Results

Cytokine profiles indicated high amounts of mRNA for interleukins 6 and 10 and transforming growth factor β. In the BAL fluid and lung parenchyma, macrophages were the predominant cell type, although the proportion was lower in the parenchyma. Lymphocytes made up approximately 3% of both cell populations. Common to both lung compartments was the predominance of CD2+ and γδ T cells over B lymphocytes. There were more CD8+ T cells than CD4+ T cells in both compartments. The γδ cells made up approximately 9% of the lymphocyte populations. Two-color flow cytometry revealed CD8+ γδ T cell and CD8+CD5 populations that were unique to BAL fluid. In the BAL fluid and parenchyma, most CD4+ and CD8+T cells expressed high amounts of CD44, a characteristic of memory T cells. The γδ T cells were CD44lo, as were B cells in the lung parenchyma. The B cells from BAL fluid expressed high amounts of CD44. Immunohistologic analysis of lung tissue revealed bronchus-associated lymphoid tissue structures with distinctive germinal center organization of B cells encompassed by CD4+ T cells.

Conclusions

Results provided normal values for comparison with those of other species and with the bovine respiratory tract response to disease. (Am J Vet Res 1997;58:969–975)

Abstract

Objective

To characterize cytokine profiles and lymphocyte subpopulations in lung parenchyma and bronchoalveolar lavage (BAL) fluid from normal bovine lungs.

Animals

Eight 12- to 18-month-old cattle.

Procedure

Cell populations in BAL fluid and collagenase-digested lung parenchyma were analyzed by flow cytometry and monoclonal antibodies. Proportions of total cell populations were determined, using Giemsa-stained cytospots. Distribution of lymphocytes within the lung parenchyma was analyzed by immunohistochemistry, and cytokine mRNA species in the parenchyma were characterized by use of reverse transcriptase-polymerase chain reaction analysis.

Results

Cytokine profiles indicated high amounts of mRNA for interleukins 6 and 10 and transforming growth factor β. In the BAL fluid and lung parenchyma, macrophages were the predominant cell type, although the proportion was lower in the parenchyma. Lymphocytes made up approximately 3% of both cell populations. Common to both lung compartments was the predominance of CD2+ and γδ T cells over B lymphocytes. There were more CD8+ T cells than CD4+ T cells in both compartments. The γδ cells made up approximately 9% of the lymphocyte populations. Two-color flow cytometry revealed CD8+ γδ T cell and CD8+CD5 populations that were unique to BAL fluid. In the BAL fluid and parenchyma, most CD4+ and CD8+T cells expressed high amounts of CD44, a characteristic of memory T cells. The γδ T cells were CD44lo, as were B cells in the lung parenchyma. The B cells from BAL fluid expressed high amounts of CD44. Immunohistologic analysis of lung tissue revealed bronchus-associated lymphoid tissue structures with distinctive germinal center organization of B cells encompassed by CD4+ T cells.

Conclusions

Results provided normal values for comparison with those of other species and with the bovine respiratory tract response to disease. (Am J Vet Res 1997;58:969–975)

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