- Email: [email protected]
Activated T Cells, but Not Lung Tumor Cells, Express Fas Ligand
Ann Thorac Surg 2014;97:378–83
Reply To the Editor: We thank Hajj-Chahine and colleagues [1] for their interest in our recent review about the mechanisms of formation of postoperative intrapericardial adhesions and their prevention [2]. Their letter gives us the opportunity to discuss an interesting article from Kuschel and colleagues [3]. We could not quote this article because it had been published electronically when our paper was already under review. After an experimental study on rabbits, Kuschel and colleagues [3] suggested that TachoSil was effective in the prevention of pericardial adhesions. In our opinion, such a conclusion deserves some comments. First, TachoSil was compared with a physical barrier, like Gore-Tex, not with a sealant, like polyethylene glycol, or with hyaluronic acid membrane. It is well known that Gore-Tex, albeit quite effective as a barrier to protect the heart, does not prevent intrapericardial adhesions [2]. Indeed, it induces a strong fibrotic reaction on epicardial surfaces. A similar finding was observed also in the study from Kuschel and colleagues [3]. Therefore, it is unsurprising that TachoSil showed superior efficacy in comparison with GoreTex for prevention of adhesions. Nevertheless, significantly fewer and less dense adhesions were observed in the retrosternal region after the application of TachoSil than in control subjects with an uncovered pericardial defect. Moreover, TachoSil is composed of equine collagen and human fibrinogen and thrombin, and the risk of anaphylactic reaction and transmission of infectious diseases cannot be understated. This is a potential disadvantage compared with other more biologically inert means to prevent adhesions, such as polyethylene glycol and hyaluronic acid. Finally, TachoSil persists at 6 weeks and disappears at 12 weeks [4], whereas hyaluronic acid and polyethylene glycol are completely resorbed within 1 and 4 weeks from application, respectively. It is unknown whether reoperations performed within a few months from the first procedure could be complicated because of the longer persistence of TachoSil. In conclusion, the encouraging results about the efficacy of TachoSil should be confirmed by comparison with other products with well-proven efficacy in the prevention of intrapericardial adhesions, such as polyethylene glycol and hyaluronic acid. Aldo Cannata, MD Luigi Martinelli, MD Department of Cardiac Surgery Niguarda Ca’ Granda Hospital P.zza Ospedale Maggiore, 3 20162 Milan, Italy e-mail: [email protected]
References 1. Hajj-Chahine J, Jayle C, Tomasi J, Corbi P. TachoSil to prevent postoperative pericardial adhesions (letter). Ann Thorac Surg 2014;97:378–9. 2. Cannata A, Petrella D, Russo CF, et al. Postsurgical intrapericardial adhesions: mechanisms of formation and prevention. Ann Thorac Surg 2013;95:1818–26. Ó 2014 by The Society of Thoracic Surgeons Published by Elsevier Inc
379
3. Kuschel TJ, Gruszka A, Hermanns-Sachweh B, et al. Prevention of postoperative pericardial adhesions with TachoSil. Ann Thorac Surg 2013;95:183–8. 4. Getman V, Devyatko E, Wolner E, Aharinejad S, Mueller MR. Fleece bound sealing prevents pleural adhesions. Interact Cardiovasc Thorac Surg 2006;5:243–6.
Correction of Anterior Leaflet Prolapse To the Editor: While reviewing an article for repair of anterior leaflet prolapse, I came across the article by Fundar o and colleagues [1]. In this article, the authors claim the technique as a personal technique used by them in 47 patients. This technique of chordal shortening at cusp level was originally published by us in 1992 [2]. In fact, the figure 1 of Fundaro’s article describes exactly the same technique we have used. We also published long-term results in 226 patients in 2004 [3]. It is quite surprising that the authors have not searched the English-language literature before claiming the technique as a personal one. The technique was first described by Frater [4] in experimental animals and later by us in our clinical experience. Arkalgud Sampathkumar, MCh Pushpanjali Crosslay Hospital W3 sector 1, Vaishali, Ghaziabad, NCR, UP 201012, India e-mail: [email protected]
References 1. Fundar o P, Moneta A, Villa E, et al. Chordal plication and free edge remodeling for mitral anterior leaflet prolapse repair: 8-year follow-up. Ann Thorac Surg 2001;72:1515–9. 2. Kumar AS, Bhan A, Kumar RV, Shrivastava S, Sood AK, Gopinath N. Cusp level chordal shortening for rheumatic mitral regurgitation. Texas Heart Inst J 1992;19:47–50. 3. Murala JSK, Kumar AS. Long-term results of cusp level chordal shortening for anterior mitral leaflet prolapse. Texas Heart Inst J 2004;31:246–50. 4. Frater RWM. Mitral valvuloplasty. In: Roberts AJ, Conti CR, eds. Current Surgery of the Heart. Philadelphia, PA: Lippincott; 1987:62–77.
Activated T Cells, but Not Lung Tumor Cells, Express Fas Ligand To the Editor : We read with great interest the recent article by Lin and colleagues [1] in which they conclude that lung cancer cells express Fas ligand (FasL) and induce death of tumor-infiltrating lymphocytes (TILs). However, reports not discussed in this study show that lung cancer cells from resected tissue, malignant pleural effusions, and cell lines do not express FasL [2, 3]. TILs rather than tumor cells express FasL [2]. Regarding the study of Lin and colleagues [1], some technical concerns should be addressed. It is unclear whether NOK-1 antibody, which is not suitable for Western blot, was used for analyzing the expression of FasL in cell lysates. Primary tumor cultures might contain FasL-expressing stroma cells, which would be responsible for bystander TILs death and reduce cytotoxic activity of TILs against tumor cells. Lin and colleagues, using a procedure previously reported by us for apoptosis detection [3], report that A549 cells induce FasL-mediated 0003-4975/$36.00
MISCELLANEOUS
surgery–a systematic review. Expert Opin Biol Ther 2009;9: 897–907. 4. Getman V, Devyatko E, Wolner E, Aharinejad S, Mueller MR. Fleece bound sealing prevents pleural adhesions. Interact Cardiovasc Thorac Surg 2006;5:243–6.
CORRESPONDENCE
380
CORRESPONDENCE
apoptosis in stimulated Jurkat cells. However, percentages of apoptotic cells are too small (<1%) to have a clinical effect. Furthermore, supernatants from A549 cells impair nuclear translocation of NF-kB that leads to apoptosis of PMA-stimulated Jurkat cells, which is a phenomenon not mediated by FasL [4]. We showed that lung cancer cells do not induce apoptosis in nonstimulated Jurkat cells that are FasL sensitive [3]. Activation-induced cell death (AICD) is a process in which Fas and FasL are upregulated after tumor recognition and T cell activation, phenomenon observed but not discussed by Lin and colleagues, leading to T cell death. AICD, rather than tumor counterattack, has been proposed to be responsible for TILs death. Recently, we reported that pleural effusion CD8þ T cells from patients with lung cancer die by AICD [5]. The tumor counterattack hypothesis is still controversial because of a lack of convincing evidence. Instead, we propose that TILs death is mediated by Fas-FasL interactions that are induced, but not mediated by lung tumors. Heriberto Prado-Garcia, PhD Susana Romero-Garcia, PhD Jose Sullivan Lopez-Gonzalez, PhD Department of Chronic-Degenerative Diseases National Institute of Respiratory Diseases Ismael Cosio Villegas Tlalpan 4502 14080 Mexico City, Mexico e-mail: [email protected]
References
MISCELLANEOUS
1. Lin Y, Liu L, Zhang T, Liu J. Functional investigation of Fas ligand expressions in human non-small cell lung cancer cells and its clinical implications. Ann Thorac Surg 2013;95:412–8. 2. Toomey D, Smyth G, Condron C, et al. Infiltrating immune cells, but not tumour cells, express FasL in non-small cell lung cancer: no association with prognosis identified in 3-year follow-up. Int J Cancer 2003;103:408–12. 3. Prado-Garcia H, Aguilar-Cazares D, Meneses-Flores M, et al. Lung carcinomas do not induce T-cell apoptosis via the Fas/ Fas ligand pathway but down-regulate CD3 epsilon expression. Cancer Immunol Immunother 2008;57:325–6. 4. Batra RK, Lin Y, Sharma S, et al. Non-small cell lung cancerderived soluble mediators enhance apoptosis in activated T lymphocytes through an I kappa B kinase-dependent mechanism. Cancer Res 2003;63:642–6. 5. Prado-Garcia H, Romero-Garcia S, Morales-Fuentes J, et al. Activation-induced cell death of memory CD8þ T cells from pleural effusion of lung cancer patients is mediated by the type II Fas-induced apoptotic pathway. Cancer Immunol Immunother 2012;61:1065–80.
Reply To the Editor: Many thanks to Dr Prado-Garcia and colleagues [1] for their interest in our publication. Generally speaking, the materials, especially the cells, we used [2] are quite different from those used by Dr Prado-Garcia’s team in their studies. The lung cancer cells they used were actually from pleural effusion [3], but we are used surgical tissue samples or cancer cells cultured from surgical tissue samples [3–5]. The cell lines we used are also different from theirs. Therefore, the results might not be comparable. But I am still pleased to discuss this matter with Dr Prado-Garcia. The expression of FasL in human lung cancer cell lines and surgical samples from patients has been proved not only by us Ó 2014 by The Society of Thoracic Surgeons Published by Elsevier Inc
Ann Thorac Surg 2014;97:378–83
but also by American [2] and European [4] researchers. Using Western blot and immunohistochemistry, we have clearly demonstrated the expression of FasL in human lung cancer [3–5]. Similar to the function of FasL, the expression of FasL might be affected by the microenvironment either within the cell or in the detecting system. There are different colons of NOK-1. NOK-1 (cat. 556387) is suitable for Western blot analysis and immunohistochemical staining. NOK-1 (cat. 556371) has been shown to neutralize the cytotoxic activity of FasL and should be used for all functional assays. Details of their uses can be found in the BD Pharmingen technical data sheet or in another authors’ publication [6]. In Figure 2B of our article, 0.1 on the Y axis represents 10% rather than 0.1% or 1%, so the difference of apoptosis is significant. We did not check the impact of the supernatant from cancer cells on T cells. We use only supernatant from target cells for the detection of radioactivity. In summary, our data demonstrate that counterattack plays an important role in fighting against T cells in the setting of lung cancer cells expressing FasL. However, as demonstrated by Dr Prado-Garcia, activation induced cell death may be the main cause of T cell death when FasL disappears from lung cancer. Yidan Lin, MD, PhD Department of Thoracic Surgery West China Hospital West China Medical School of Sichuan University 37 Guo Xue Alley Chengdu, Sichuan Province, China 610041 e-mail: [email protected]
References 1. Prado-Garcia H, Romero-Garcia S, Lopez-Gonzalez JS. Activated T cells, but not lung tumor cells, express Fas ligand (letter). Ann Thorac Surg 2014;97:379–80. 2. Niehans GA, Brunner T, Frizelle SP, et al. Human lung carcinomas express Fas ligand. Cancer Res 1997;57: 1007–12. 3. Prado-Garcia H, Aguilar-Cazares D, Meneses-Flores M, Morales-Fuentes J, Lopez-Gonzalez JS. Lung carcinomas do not induce T-cell apoptosis via the Fas/Fas ligand pathway but down-regulate CD3 epsilon expression. Cancer Immunol Immunother 2008;57:325–36. 4. Koomagi R, Volm M. Expression of Fas (CD95/APO-1) and Fas ligand in lung cancer, its prognostic and predictive relevance. Int J Cancer 1999;84:239–43. 5. Lin Y, Liu L, Zhang T, Liu J. Functional investigation of Fas ligand expressions in human non-small cell lung cancer cells and its clinical implications. Ann Thorac Surg 2013;95: 412–8. 6. Takahashi T, Tanaka M, Brannan CI, et al. Generalized lymphoproliferative disease in mice, caused by a point mutation in the Fas ligand. Cell 1994;l76:969–76.
Immunohistochemical Improvement in the Analysis of the Lymphatic Metastases From Lung Carcinoma To the Editor: We read with great interest the article by Strano et al [1] concerning with the prognostic significance of vascular and lymphatic metastases from non-small-cell lung carcinoma. It is well known that it is not morphologically possible to distinguish vascular and lymphatic capillaries without a clear evidence of erythrocytes inside the blood capillary. Moreover, the 0003-4975/$36.00
Reply To the Editor: We thank Hajj-Chahine and colleagues [1] for their interest in our recent review about the mechanisms of formation of postoperative intrapericardial adhesions and their prevention [2]. Their letter gives us the opportunity to discuss an interesting article from Kuschel and colleagues [3]. We could not quote this article because it had been published electronically when our paper was already under review. After an experimental study on rabbits, Kuschel and colleagues [3] suggested that TachoSil was effective in the prevention of pericardial adhesions. In our opinion, such a conclusion deserves some comments. First, TachoSil was compared with a physical barrier, like Gore-Tex, not with a sealant, like polyethylene glycol, or with hyaluronic acid membrane. It is well known that Gore-Tex, albeit quite effective as a barrier to protect the heart, does not prevent intrapericardial adhesions [2]. Indeed, it induces a strong fibrotic reaction on epicardial surfaces. A similar finding was observed also in the study from Kuschel and colleagues [3]. Therefore, it is unsurprising that TachoSil showed superior efficacy in comparison with GoreTex for prevention of adhesions. Nevertheless, significantly fewer and less dense adhesions were observed in the retrosternal region after the application of TachoSil than in control subjects with an uncovered pericardial defect. Moreover, TachoSil is composed of equine collagen and human fibrinogen and thrombin, and the risk of anaphylactic reaction and transmission of infectious diseases cannot be understated. This is a potential disadvantage compared with other more biologically inert means to prevent adhesions, such as polyethylene glycol and hyaluronic acid. Finally, TachoSil persists at 6 weeks and disappears at 12 weeks [4], whereas hyaluronic acid and polyethylene glycol are completely resorbed within 1 and 4 weeks from application, respectively. It is unknown whether reoperations performed within a few months from the first procedure could be complicated because of the longer persistence of TachoSil. In conclusion, the encouraging results about the efficacy of TachoSil should be confirmed by comparison with other products with well-proven efficacy in the prevention of intrapericardial adhesions, such as polyethylene glycol and hyaluronic acid. Aldo Cannata, MD Luigi Martinelli, MD Department of Cardiac Surgery Niguarda Ca’ Granda Hospital P.zza Ospedale Maggiore, 3 20162 Milan, Italy e-mail: [email protected]
References 1. Hajj-Chahine J, Jayle C, Tomasi J, Corbi P. TachoSil to prevent postoperative pericardial adhesions (letter). Ann Thorac Surg 2014;97:378–9. 2. Cannata A, Petrella D, Russo CF, et al. Postsurgical intrapericardial adhesions: mechanisms of formation and prevention. Ann Thorac Surg 2013;95:1818–26. Ó 2014 by The Society of Thoracic Surgeons Published by Elsevier Inc
379
3. Kuschel TJ, Gruszka A, Hermanns-Sachweh B, et al. Prevention of postoperative pericardial adhesions with TachoSil. Ann Thorac Surg 2013;95:183–8. 4. Getman V, Devyatko E, Wolner E, Aharinejad S, Mueller MR. Fleece bound sealing prevents pleural adhesions. Interact Cardiovasc Thorac Surg 2006;5:243–6.
Correction of Anterior Leaflet Prolapse To the Editor: While reviewing an article for repair of anterior leaflet prolapse, I came across the article by Fundar o and colleagues [1]. In this article, the authors claim the technique as a personal technique used by them in 47 patients. This technique of chordal shortening at cusp level was originally published by us in 1992 [2]. In fact, the figure 1 of Fundaro’s article describes exactly the same technique we have used. We also published long-term results in 226 patients in 2004 [3]. It is quite surprising that the authors have not searched the English-language literature before claiming the technique as a personal one. The technique was first described by Frater [4] in experimental animals and later by us in our clinical experience. Arkalgud Sampathkumar, MCh Pushpanjali Crosslay Hospital W3 sector 1, Vaishali, Ghaziabad, NCR, UP 201012, India e-mail: [email protected]
References 1. Fundar o P, Moneta A, Villa E, et al. Chordal plication and free edge remodeling for mitral anterior leaflet prolapse repair: 8-year follow-up. Ann Thorac Surg 2001;72:1515–9. 2. Kumar AS, Bhan A, Kumar RV, Shrivastava S, Sood AK, Gopinath N. Cusp level chordal shortening for rheumatic mitral regurgitation. Texas Heart Inst J 1992;19:47–50. 3. Murala JSK, Kumar AS. Long-term results of cusp level chordal shortening for anterior mitral leaflet prolapse. Texas Heart Inst J 2004;31:246–50. 4. Frater RWM. Mitral valvuloplasty. In: Roberts AJ, Conti CR, eds. Current Surgery of the Heart. Philadelphia, PA: Lippincott; 1987:62–77.
Activated T Cells, but Not Lung Tumor Cells, Express Fas Ligand To the Editor : We read with great interest the recent article by Lin and colleagues [1] in which they conclude that lung cancer cells express Fas ligand (FasL) and induce death of tumor-infiltrating lymphocytes (TILs). However, reports not discussed in this study show that lung cancer cells from resected tissue, malignant pleural effusions, and cell lines do not express FasL [2, 3]. TILs rather than tumor cells express FasL [2]. Regarding the study of Lin and colleagues [1], some technical concerns should be addressed. It is unclear whether NOK-1 antibody, which is not suitable for Western blot, was used for analyzing the expression of FasL in cell lysates. Primary tumor cultures might contain FasL-expressing stroma cells, which would be responsible for bystander TILs death and reduce cytotoxic activity of TILs against tumor cells. Lin and colleagues, using a procedure previously reported by us for apoptosis detection [3], report that A549 cells induce FasL-mediated 0003-4975/$36.00
MISCELLANEOUS
surgery–a systematic review. Expert Opin Biol Ther 2009;9: 897–907. 4. Getman V, Devyatko E, Wolner E, Aharinejad S, Mueller MR. Fleece bound sealing prevents pleural adhesions. Interact Cardiovasc Thorac Surg 2006;5:243–6.
CORRESPONDENCE
380
CORRESPONDENCE
apoptosis in stimulated Jurkat cells. However, percentages of apoptotic cells are too small (<1%) to have a clinical effect. Furthermore, supernatants from A549 cells impair nuclear translocation of NF-kB that leads to apoptosis of PMA-stimulated Jurkat cells, which is a phenomenon not mediated by FasL [4]. We showed that lung cancer cells do not induce apoptosis in nonstimulated Jurkat cells that are FasL sensitive [3]. Activation-induced cell death (AICD) is a process in which Fas and FasL are upregulated after tumor recognition and T cell activation, phenomenon observed but not discussed by Lin and colleagues, leading to T cell death. AICD, rather than tumor counterattack, has been proposed to be responsible for TILs death. Recently, we reported that pleural effusion CD8þ T cells from patients with lung cancer die by AICD [5]. The tumor counterattack hypothesis is still controversial because of a lack of convincing evidence. Instead, we propose that TILs death is mediated by Fas-FasL interactions that are induced, but not mediated by lung tumors. Heriberto Prado-Garcia, PhD Susana Romero-Garcia, PhD Jose Sullivan Lopez-Gonzalez, PhD Department of Chronic-Degenerative Diseases National Institute of Respiratory Diseases Ismael Cosio Villegas Tlalpan 4502 14080 Mexico City, Mexico e-mail: [email protected]
References
MISCELLANEOUS
1. Lin Y, Liu L, Zhang T, Liu J. Functional investigation of Fas ligand expressions in human non-small cell lung cancer cells and its clinical implications. Ann Thorac Surg 2013;95:412–8. 2. Toomey D, Smyth G, Condron C, et al. Infiltrating immune cells, but not tumour cells, express FasL in non-small cell lung cancer: no association with prognosis identified in 3-year follow-up. Int J Cancer 2003;103:408–12. 3. Prado-Garcia H, Aguilar-Cazares D, Meneses-Flores M, et al. Lung carcinomas do not induce T-cell apoptosis via the Fas/ Fas ligand pathway but down-regulate CD3 epsilon expression. Cancer Immunol Immunother 2008;57:325–6. 4. Batra RK, Lin Y, Sharma S, et al. Non-small cell lung cancerderived soluble mediators enhance apoptosis in activated T lymphocytes through an I kappa B kinase-dependent mechanism. Cancer Res 2003;63:642–6. 5. Prado-Garcia H, Romero-Garcia S, Morales-Fuentes J, et al. Activation-induced cell death of memory CD8þ T cells from pleural effusion of lung cancer patients is mediated by the type II Fas-induced apoptotic pathway. Cancer Immunol Immunother 2012;61:1065–80.
Reply To the Editor: Many thanks to Dr Prado-Garcia and colleagues [1] for their interest in our publication. Generally speaking, the materials, especially the cells, we used [2] are quite different from those used by Dr Prado-Garcia’s team in their studies. The lung cancer cells they used were actually from pleural effusion [3], but we are used surgical tissue samples or cancer cells cultured from surgical tissue samples [3–5]. The cell lines we used are also different from theirs. Therefore, the results might not be comparable. But I am still pleased to discuss this matter with Dr Prado-Garcia. The expression of FasL in human lung cancer cell lines and surgical samples from patients has been proved not only by us Ó 2014 by The Society of Thoracic Surgeons Published by Elsevier Inc
Ann Thorac Surg 2014;97:378–83
but also by American [2] and European [4] researchers. Using Western blot and immunohistochemistry, we have clearly demonstrated the expression of FasL in human lung cancer [3–5]. Similar to the function of FasL, the expression of FasL might be affected by the microenvironment either within the cell or in the detecting system. There are different colons of NOK-1. NOK-1 (cat. 556387) is suitable for Western blot analysis and immunohistochemical staining. NOK-1 (cat. 556371) has been shown to neutralize the cytotoxic activity of FasL and should be used for all functional assays. Details of their uses can be found in the BD Pharmingen technical data sheet or in another authors’ publication [6]. In Figure 2B of our article, 0.1 on the Y axis represents 10% rather than 0.1% or 1%, so the difference of apoptosis is significant. We did not check the impact of the supernatant from cancer cells on T cells. We use only supernatant from target cells for the detection of radioactivity. In summary, our data demonstrate that counterattack plays an important role in fighting against T cells in the setting of lung cancer cells expressing FasL. However, as demonstrated by Dr Prado-Garcia, activation induced cell death may be the main cause of T cell death when FasL disappears from lung cancer. Yidan Lin, MD, PhD Department of Thoracic Surgery West China Hospital West China Medical School of Sichuan University 37 Guo Xue Alley Chengdu, Sichuan Province, China 610041 e-mail: [email protected]
References 1. Prado-Garcia H, Romero-Garcia S, Lopez-Gonzalez JS. Activated T cells, but not lung tumor cells, express Fas ligand (letter). Ann Thorac Surg 2014;97:379–80. 2. Niehans GA, Brunner T, Frizelle SP, et al. Human lung carcinomas express Fas ligand. Cancer Res 1997;57: 1007–12. 3. Prado-Garcia H, Aguilar-Cazares D, Meneses-Flores M, Morales-Fuentes J, Lopez-Gonzalez JS. Lung carcinomas do not induce T-cell apoptosis via the Fas/Fas ligand pathway but down-regulate CD3 epsilon expression. Cancer Immunol Immunother 2008;57:325–36. 4. Koomagi R, Volm M. Expression of Fas (CD95/APO-1) and Fas ligand in lung cancer, its prognostic and predictive relevance. Int J Cancer 1999;84:239–43. 5. Lin Y, Liu L, Zhang T, Liu J. Functional investigation of Fas ligand expressions in human non-small cell lung cancer cells and its clinical implications. Ann Thorac Surg 2013;95: 412–8. 6. Takahashi T, Tanaka M, Brannan CI, et al. Generalized lymphoproliferative disease in mice, caused by a point mutation in the Fas ligand. Cell 1994;l76:969–76.
Immunohistochemical Improvement in the Analysis of the Lymphatic Metastases From Lung Carcinoma To the Editor: We read with great interest the article by Strano et al [1] concerning with the prognostic significance of vascular and lymphatic metastases from non-small-cell lung carcinoma. It is well known that it is not morphologically possible to distinguish vascular and lymphatic capillaries without a clear evidence of erythrocytes inside the blood capillary. Moreover, the 0003-4975/$36.00