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Alveolar Macrophage-Lymphocyte Interaction
Alveolar Macrophage-lymphocyte lnteradion* Alterations in Smokers and in Sarcoidosis Henty Yeuger, Jr., M.D.,F.C.C.P.; L. W , B.S.; and Jag Prashad, M.D.
and lymphocytes must interact physiMacrophages cally in the generation of many cell-mediated im-
mune reactions in oitro, and presumably in oioo. We performed studies to evaluate the capabilities of alveolar macrophages (AM) to interact with autologous blood lymphocytes .(L) in two settings in which AM-L interaction might be expected to be altered, in cigarette smokers, and in pelrons with sarcoidosis. Lymphocytes were separated by Ficoll-Hypaque sedimentation and nylon wool column pdcation. AM obtained by saline bronchial lavage were allowed to attach in monolayers in Lab-Tek chambers. Lymphocytes were added and the cells were allowed to incubate varying intervals of time. AM:L binding was assessed visually. Cells fnnn six normal non-smokers, four normal smokers, four smdrem with sarcoidosis, and two non smdcers with sarcoidosis have been evaluated. In the absence of added antigen, cells from smokers showed decreased binding, and nonsmoking patients with sarcoid showed increased binding at 2 and 18 hours. Cells from patients with sarcoidosis who smoked responded like other smokers' cells. In the presence of Kveim suspension, AM-L interaction was partially inhibited in six o f six samid patients; spleen suspension had no effect. In the two sarcoid patients with positive skin tests to recall antigens, AM-L binding was increased in the presence of the recall antigen; control subjects showed a similar degree of increased AM-L binding in the presence of recall antigens to which they had positive skin tests. Thus, smokers' alveolar macrophages have decreased, and nonsmoking sarcoidosispatients' AM show increased L binding in the absence of antigen; the functional implications are not clear. In addition, this system shows promise for looking at mechanisms of Kveim effect on sarcoid mononuclear cells; Kveim suspension appears to have effects Werent from conventional recan antigens.
*From the Pulmonary Disease Division and Immunology Center, Georgetown Medical Center and District of Columbia General H ital, Washington, D.C. s u p p o d by
ZPHS w t HLB ~m etacn Untostsfty HospG
Reprfnt mquests: Dr. Yeager,
td,3800 Reservoir Rwd NW,%%fngton,
DC 20007
Dr. Mitchell: Do you think your findings in smokers might help explain their tendency to have increased incidence of respiratory infections?
CHEST, 75: 2, FEBRUARY, 1979 SUf'PlEMENT
Dr. Yeuger: Yes. There are a number of functional abnormalities in smokers' cells that probably contribute to the tendency of smokers to have more infections. Question: Did you control the ratio of lymphocytes to macrophages in your mixtures? Dr. Yeager: Yes. In most of our data there was a standard lymphocyte to macrophage ratio of 50:l. Qwstiun: Did you try varying this ratio to see if that might make a difference? Dr. Yeager: Not in the sarcoid patients, where the standard ratio was 50:l. In normal subjects we have tried variation, and it appears to affect the results in a dose-dependent fashion. Question: In order for lymphocytes and macrophages to interact to augment response to a given antigen, they must share certain histocompatibility antigens. Have you tried reacting macrophages with peripheral blood lymphocytes from genetically dissimilar individuals to see if these antigens are participating in the interaction? Dr. Yeager: What you say is true and this is why so far we have stuck with autologous lymphocytes. Dr. Turner-Warwick:I wonder if using lymphocytes fnmn lavage fluid of sarcoid patients would have affected the response to the Kveim antigen? Dr. Yeuger: We have not yet done that. Dr. Davis: Are the lymphocytes in the macrophage r m e s T cells or B cells? Dr. Yeager: We suspect that they are mostly T cells, but have not systematically looked at that question yet. Dr. Sunderson: It's possible that lymph could be decreased in peripheral blood because of involvement in the sarcoid reaction. What do you think? Dr. Yeager: I think that's possible. Dr. Tdmage: Did you try pre-heating the lymphocytes to see if heat-killed lymph bound differently to macrophages? L)r. Yeuger: We have not tried that in the sarcoid patients. In normal subjects, heat-killing of lymphocytes seemed not to alter binding of lymphocytes to alveolar macrophages, or at times to enhance it slightly. Dr. Shasby: Did you try to incubate lymphs from sarcoid patients in sarcoid serum? Dr. Yecrger:We have not yet done that. Dr. Rylander: Any signs of reduced macrophage activity in the macrophages of sarcoidosispatients? Dr. Yeager: We have not done detailed metabolic or functional studies on these ceIls. Dr. Shusby: It would be of interest to see if NO, would interfere with the immune reaction of macrophages. Dt. Fruhmunn: Do the people here feel sarcoid is primarily an alveolitis or a reaction of the connective tissue at least in the more commonly seen early stages? Dr. Yeager: I feel that there is evidence for both interstitial and intra-alveolar inflammation, and at this stage we can't say much more than that.
IMMUWOLO6Y OF THE LUW6 288
and lymphocytes must interact physiMacrophages cally in the generation of many cell-mediated im-
mune reactions in oitro, and presumably in oioo. We performed studies to evaluate the capabilities of alveolar macrophages (AM) to interact with autologous blood lymphocytes .(L) in two settings in which AM-L interaction might be expected to be altered, in cigarette smokers, and in pelrons with sarcoidosis. Lymphocytes were separated by Ficoll-Hypaque sedimentation and nylon wool column pdcation. AM obtained by saline bronchial lavage were allowed to attach in monolayers in Lab-Tek chambers. Lymphocytes were added and the cells were allowed to incubate varying intervals of time. AM:L binding was assessed visually. Cells fnnn six normal non-smokers, four normal smokers, four smdrem with sarcoidosis, and two non smdcers with sarcoidosis have been evaluated. In the absence of added antigen, cells from smokers showed decreased binding, and nonsmoking patients with sarcoid showed increased binding at 2 and 18 hours. Cells from patients with sarcoidosis who smoked responded like other smokers' cells. In the presence of Kveim suspension, AM-L interaction was partially inhibited in six o f six samid patients; spleen suspension had no effect. In the two sarcoid patients with positive skin tests to recall antigens, AM-L binding was increased in the presence of the recall antigen; control subjects showed a similar degree of increased AM-L binding in the presence of recall antigens to which they had positive skin tests. Thus, smokers' alveolar macrophages have decreased, and nonsmoking sarcoidosispatients' AM show increased L binding in the absence of antigen; the functional implications are not clear. In addition, this system shows promise for looking at mechanisms of Kveim effect on sarcoid mononuclear cells; Kveim suspension appears to have effects Werent from conventional recan antigens.
*From the Pulmonary Disease Division and Immunology Center, Georgetown Medical Center and District of Columbia General H ital, Washington, D.C. s u p p o d by
ZPHS w t HLB ~m etacn Untostsfty HospG
Reprfnt mquests: Dr. Yeager,
td,3800 Reservoir Rwd NW,%%fngton,
DC 20007
Dr. Mitchell: Do you think your findings in smokers might help explain their tendency to have increased incidence of respiratory infections?
CHEST, 75: 2, FEBRUARY, 1979 SUf'PlEMENT
Dr. Yeuger: Yes. There are a number of functional abnormalities in smokers' cells that probably contribute to the tendency of smokers to have more infections. Question: Did you control the ratio of lymphocytes to macrophages in your mixtures? Dr. Yeager: Yes. In most of our data there was a standard lymphocyte to macrophage ratio of 50:l. Qwstiun: Did you try varying this ratio to see if that might make a difference? Dr. Yeager: Not in the sarcoid patients, where the standard ratio was 50:l. In normal subjects we have tried variation, and it appears to affect the results in a dose-dependent fashion. Question: In order for lymphocytes and macrophages to interact to augment response to a given antigen, they must share certain histocompatibility antigens. Have you tried reacting macrophages with peripheral blood lymphocytes from genetically dissimilar individuals to see if these antigens are participating in the interaction? Dr. Yeager: What you say is true and this is why so far we have stuck with autologous lymphocytes. Dr. Turner-Warwick:I wonder if using lymphocytes fnmn lavage fluid of sarcoid patients would have affected the response to the Kveim antigen? Dr. Yeuger: We have not yet done that. Dr. Davis: Are the lymphocytes in the macrophage r m e s T cells or B cells? Dr. Yeager: We suspect that they are mostly T cells, but have not systematically looked at that question yet. Dr. Sunderson: It's possible that lymph could be decreased in peripheral blood because of involvement in the sarcoid reaction. What do you think? Dr. Yeager: I think that's possible. Dr. Tdmage: Did you try pre-heating the lymphocytes to see if heat-killed lymph bound differently to macrophages? L)r. Yeuger: We have not tried that in the sarcoid patients. In normal subjects, heat-killing of lymphocytes seemed not to alter binding of lymphocytes to alveolar macrophages, or at times to enhance it slightly. Dr. Shasby: Did you try to incubate lymphs from sarcoid patients in sarcoid serum? Dr. Yecrger:We have not yet done that. Dr. Rylander: Any signs of reduced macrophage activity in the macrophages of sarcoidosispatients? Dr. Yeager: We have not done detailed metabolic or functional studies on these ceIls. Dr. Shusby: It would be of interest to see if NO, would interfere with the immune reaction of macrophages. Dt. Fruhmunn: Do the people here feel sarcoid is primarily an alveolitis or a reaction of the connective tissue at least in the more commonly seen early stages? Dr. Yeager: I feel that there is evidence for both interstitial and intra-alveolar inflammation, and at this stage we can't say much more than that.
IMMUWOLO6Y OF THE LUW6 288