LETTER TO THE EDITOR Transient inhibition of neutrophil migration following plasma or plasma-platelet apheresis donation procedures Massimo Ghio1, Maria Bertolotto1, Luciano Ottonello1, Paola Contini1, Gianluca Ubezio1, Gino Tripodi2 Department of Internal Medicine, I.R.C.C.S. "A.O.U. San Martino-IST" and University of Genoa; Immunohematology and Transfusion Centre, Giannina Gaslini" Institute, Genoa, Italy
M e d i c i n e a n d Immunohaematology (SIMTI) recommendations for apheresis donation. Informed consent was obtained from all the donors. Samples were obtained from each donor to perform a complete preprocedure laboratory profile. In addition to biochemical and serological tests required by law, C-reactive protein, erythrocyte sedimentation rate, protein electrophoresis, fibrinogen and ferritin were also evaluated in order to further exclude any sub-clinical inflammatory disorder possibly causing leucocyte activation. Follow-up assessments were performed every 6 months up to 2 years, by physical examination, anamnestic evaluation and the same biochemical/serological tests done during the enrolment, as previously described1,3. Monoclonal antibodies and other reagents for research use were purchased from several renowned companies. The modified Boyden chamber migration assay was conducted using human neutrophils from different healthy volunteers isolated from heparinised venous blood and the cells were pre-incubated with chosen dilutions of plasma from donors (plasma dilution range from 1:1×104 to 1:1×105 , data not shown). Cells were washed and neutrophil migration toward control medium or human recombinant CXCL8 was assessed in a 48-well micro-chemotaxis chamber as previously described4,5. Data are expressed as net migration. The previous studies focused on sHLA-I-mediated immunomodulation following plasma-platelet apheresis donation procedures1,3, regardless of the type of donation procedure. The comparison between the different timings showed a noteworthy inhibition of neutrophil locomotion induced by plasma pre-treatment (also in comparison with matched non-pretreated controls, data not shown) which was consistently observed immediately the after plasma-platelet apheresis donation procedure. An evident and stable reversion to baseline levels was detectable 7 and 14 days after the procedure (Figure 1). In accordance with such short-lived immunomodulation, no enrolled subject experienced any adverse reaction during the procedures or showed any possible immunosuppression or viral, bacterial or neoplastic disease during 24 months of monitoring following the donations.
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Dear Sir, Our group has long been interested in the immunomodulatory role of soluble HLA class I molecules (sHLA-I) in all clinically available blood products and derivatives and recently we evaluated possible immunomodulatory effects following plasmaplatelet apheresis donation procedures. We found that, following centrifugation-based apheresis, whole and/or re-folded sHLA-I bind to the apheresis circuit surfaces fabricated from synthetic polymers during therapeutic and donation procedures1-3. Monocytes and neutrophils, or CD8+ T lymphocytes and natural killer cells could bind sHLA-I molecules with immunoglobulin-like transcripts and CD8 membrane molecules, respectively. Accordingly, we showed that all these leucocytes, rolling into the circuits during the procedure, could bind sHLA-I molecules adsorbed to the circuits' polymers and thus become sensitive to the biological effects of sHLA-I, such as transcriptional/ post-transcriptional modulation transforming growth factor-beta 1 (TGFβ1)1-3. Finally, we have previously demonstrated that high dilutions of supernatants from stored red blood cells inhibit neutrophil migration. Such inhibitory activity was demonstrated to be due to the TGFβ1 contained in the supernatants, which is capable of desensitizing neutrophils to chemotactic stimulation4. On this basis, it could be hypothesised that TGFβ1 modulation induced by plasma-platelet apheresis donation procedures may also inhibit neutrophil migration. To determine whether this is the case, samples of plasma were collected from three male donors before, and immediately, 7 and 14 days after the plasma and platelet donation procedure. Moreover, to highlight a potential unique mechanism in neutrophil locomotion inhibition following apheresis donation, we compared different plasma and/or platelet donation procedures and separators (Haemonetics PCS2, model 6002, Haemonetics MCS Plus version C [Haemonetics Corp., Baintree, MA, USA] and Trima Accel version 4.0 [Gambro BTC, Zaventem, Belgium])1,3. Apheresis donor procedures were performed in accordance with the Italian law, in healthy donors fulfilling the criteria laid down by the Italian blood donation guidelines and Italian Society of Transfusion
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Blood Transfus 2015; 13: 682-3 DOI 10.2450/2014.0257-14 © SIMTI Servizi Srl
682 All rights reserved - For personal use only No other uses without permission
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Donor apheresis procedures ephemerally inhibit neutrophils chemotaxis
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Figure 1 - Immediately after plasma-platelet apheresis, the locomotion of neutrophils from healthy subjects was constantly induced by pre-treatment donor's plasma regardless of the type of donation procedure. An evident and stable reversion to baseline levels was detectable 7 and 14 days after the procedure.
er v
The effects of Haemonetics PCS2, model 6002 plasma collection (○), Haemonetics MCS Plus version C (□) and Trima Accel version 4.0 (∆) combined plasma and platelet collection on neutrophil migration are shown.
3) 4)
Ghio M, Contini P, Ansaldi F, et al. Donor neutrophil activation and transforming growth factor-ß1 modulation induced by donor apheresis procedures. Blood Transfus 2014; 12: 615-7. Ghio M, Ottonello L, Contini P, et al. Transforming growth factor-beta1 in supernatants from stored red blood cells inhibits neutrophil locomotion. Blood 2003; 102: 1100-7. Montecucco F, Bertolotto M, Vuilleumier N, et al. Acipimox reduces circulating levels of insulin and associated neutrophilic inflammation in metabolic syndrome. Am J Physiol Endocrinol Metab 2011; 300: E681-90.
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On the whole, our findings indicate that durable inhibition of neutrophil locomotion does not occur in healthy donors undergoing plasma-platelet apheresis donation. Completing previous studies focused on sHLA-I mediated immunomodulation following plasma-platelet apheresis donation procedures, this fact might further corroborate a conceivable "immunological" harmlessness of such procedures.
5)
References
Ghio M, Contini P, Ansaldi F, et al. Immunomodulation due to plasma or plasma-platelet apheresis donation: events occurring during donation procedures. J Clin Apher 2014; DOI: 10.1002/ jca.21362 [Epub ahead of print]. Ghio M, Contini P, Ansaldi F, et al. A possible role of soluble HLA-I molecule in the immunomodulatory effects of therapeutic apheresis. Blood Transfus 2014; 12 (Suppl 1): s167-9.
©
1)
SI
The Authors declare no conflict of interest.
2)
Arrived: 13 October 2014 - Revision accepted: 6 November 2014 Correspondence: Massimo Ghio Department of Internal Medicine (DiMI) and Medical Specialties I.R.C.C.S. "A.O.U. San Martino-IST" and University of Genoa Viale Benedetto XV, 6 16132 Genoa, Italy e-mail: [email protected]
Blood Transfus 2015; 13: 682-3 DOI 10.2450/2014.0257-14 683 All rights reserved - For personal use only No other uses without permission
M e d i c i n e a n d Immunohaematology (SIMTI) recommendations for apheresis donation. Informed consent was obtained from all the donors. Samples were obtained from each donor to perform a complete preprocedure laboratory profile. In addition to biochemical and serological tests required by law, C-reactive protein, erythrocyte sedimentation rate, protein electrophoresis, fibrinogen and ferritin were also evaluated in order to further exclude any sub-clinical inflammatory disorder possibly causing leucocyte activation. Follow-up assessments were performed every 6 months up to 2 years, by physical examination, anamnestic evaluation and the same biochemical/serological tests done during the enrolment, as previously described1,3. Monoclonal antibodies and other reagents for research use were purchased from several renowned companies. The modified Boyden chamber migration assay was conducted using human neutrophils from different healthy volunteers isolated from heparinised venous blood and the cells were pre-incubated with chosen dilutions of plasma from donors (plasma dilution range from 1:1×104 to 1:1×105 , data not shown). Cells were washed and neutrophil migration toward control medium or human recombinant CXCL8 was assessed in a 48-well micro-chemotaxis chamber as previously described4,5. Data are expressed as net migration. The previous studies focused on sHLA-I-mediated immunomodulation following plasma-platelet apheresis donation procedures1,3, regardless of the type of donation procedure. The comparison between the different timings showed a noteworthy inhibition of neutrophil locomotion induced by plasma pre-treatment (also in comparison with matched non-pretreated controls, data not shown) which was consistently observed immediately the after plasma-platelet apheresis donation procedure. An evident and stable reversion to baseline levels was detectable 7 and 14 days after the procedure (Figure 1). In accordance with such short-lived immunomodulation, no enrolled subject experienced any adverse reaction during the procedures or showed any possible immunosuppression or viral, bacterial or neoplastic disease during 24 months of monitoring following the donations.
©
SI
M
TI S
er v
Dear Sir, Our group has long been interested in the immunomodulatory role of soluble HLA class I molecules (sHLA-I) in all clinically available blood products and derivatives and recently we evaluated possible immunomodulatory effects following plasmaplatelet apheresis donation procedures. We found that, following centrifugation-based apheresis, whole and/or re-folded sHLA-I bind to the apheresis circuit surfaces fabricated from synthetic polymers during therapeutic and donation procedures1-3. Monocytes and neutrophils, or CD8+ T lymphocytes and natural killer cells could bind sHLA-I molecules with immunoglobulin-like transcripts and CD8 membrane molecules, respectively. Accordingly, we showed that all these leucocytes, rolling into the circuits during the procedure, could bind sHLA-I molecules adsorbed to the circuits' polymers and thus become sensitive to the biological effects of sHLA-I, such as transcriptional/ post-transcriptional modulation transforming growth factor-beta 1 (TGFβ1)1-3. Finally, we have previously demonstrated that high dilutions of supernatants from stored red blood cells inhibit neutrophil migration. Such inhibitory activity was demonstrated to be due to the TGFβ1 contained in the supernatants, which is capable of desensitizing neutrophils to chemotactic stimulation4. On this basis, it could be hypothesised that TGFβ1 modulation induced by plasma-platelet apheresis donation procedures may also inhibit neutrophil migration. To determine whether this is the case, samples of plasma were collected from three male donors before, and immediately, 7 and 14 days after the plasma and platelet donation procedure. Moreover, to highlight a potential unique mechanism in neutrophil locomotion inhibition following apheresis donation, we compared different plasma and/or platelet donation procedures and separators (Haemonetics PCS2, model 6002, Haemonetics MCS Plus version C [Haemonetics Corp., Baintree, MA, USA] and Trima Accel version 4.0 [Gambro BTC, Zaventem, Belgium])1,3. Apheresis donor procedures were performed in accordance with the Italian law, in healthy donors fulfilling the criteria laid down by the Italian blood donation guidelines and Italian Society of Transfusion
Sr l
2
iz i
1
Blood Transfus 2015; 13: 682-3 DOI 10.2450/2014.0257-14 © SIMTI Servizi Srl
682 All rights reserved - For personal use only No other uses without permission
Sr l
Donor apheresis procedures ephemerally inhibit neutrophils chemotaxis
iz i
Figure 1 - Immediately after plasma-platelet apheresis, the locomotion of neutrophils from healthy subjects was constantly induced by pre-treatment donor's plasma regardless of the type of donation procedure. An evident and stable reversion to baseline levels was detectable 7 and 14 days after the procedure.
er v
The effects of Haemonetics PCS2, model 6002 plasma collection (○), Haemonetics MCS Plus version C (□) and Trima Accel version 4.0 (∆) combined plasma and platelet collection on neutrophil migration are shown.
3) 4)
Ghio M, Contini P, Ansaldi F, et al. Donor neutrophil activation and transforming growth factor-ß1 modulation induced by donor apheresis procedures. Blood Transfus 2014; 12: 615-7. Ghio M, Ottonello L, Contini P, et al. Transforming growth factor-beta1 in supernatants from stored red blood cells inhibits neutrophil locomotion. Blood 2003; 102: 1100-7. Montecucco F, Bertolotto M, Vuilleumier N, et al. Acipimox reduces circulating levels of insulin and associated neutrophilic inflammation in metabolic syndrome. Am J Physiol Endocrinol Metab 2011; 300: E681-90.
M
TI S
On the whole, our findings indicate that durable inhibition of neutrophil locomotion does not occur in healthy donors undergoing plasma-platelet apheresis donation. Completing previous studies focused on sHLA-I mediated immunomodulation following plasma-platelet apheresis donation procedures, this fact might further corroborate a conceivable "immunological" harmlessness of such procedures.
5)
References
Ghio M, Contini P, Ansaldi F, et al. Immunomodulation due to plasma or plasma-platelet apheresis donation: events occurring during donation procedures. J Clin Apher 2014; DOI: 10.1002/ jca.21362 [Epub ahead of print]. Ghio M, Contini P, Ansaldi F, et al. A possible role of soluble HLA-I molecule in the immunomodulatory effects of therapeutic apheresis. Blood Transfus 2014; 12 (Suppl 1): s167-9.
©
1)
SI
The Authors declare no conflict of interest.
2)
Arrived: 13 October 2014 - Revision accepted: 6 November 2014 Correspondence: Massimo Ghio Department of Internal Medicine (DiMI) and Medical Specialties I.R.C.C.S. "A.O.U. San Martino-IST" and University of Genoa Viale Benedetto XV, 6 16132 Genoa, Italy e-mail: [email protected]
Blood Transfus 2015; 13: 682-3 DOI 10.2450/2014.0257-14 683 All rights reserved - For personal use only No other uses without permission
