neutrophil activation in immunoadsorption

Neutrophil Activation in Immunoadsorption

*Kohei Ota, *Yuko Shimizu, *Hisae Ichikawa, *Mika Ueda, *Naoko Akiyama,†Chieko Higuchi, †Tetsuzo Agishi, and *Makoto Iwata

*Department of Neurology, Neurological Institute, and †Division of Blood Purification, Kidney Center, Tokyo Women’sMedical University, Tokyo, Japan

Abstract: Immunoadsorption therapy (IAT) is used in thetreatment of autoimmune diseases. Although IAT hasbeen reported to modify humoral immunity by inducingchemokines and activating complements, much remainsunknown about the biological effects of IAT on cellularcomponents in peripheral blood. To define the influenceof IAT on leukocytes, we determined leukocyte L-selectin(CD62L) and Mac-1 (CD11b) as parameters for activationof leukocytes in peripheral blood during IAT. Peripheralleukocyte L-selectin and Mac-1 were determined continu-ously by flow cytometry in 6 patients with neuroimmuno-logical disorders in whom IAT was conducted usinga Plasma Flow OP-05 (Asahi Medical Corp., Tokyo, Japan)as a plasma separator and Immusorba TR-350 (AsahiMedical Corp., Tokyo, Japan) as an adsorption column.Expression of neutrophils (PMN) L-selectin was decreased

30 min after starting IAT, with the decreases particularlymarked at the end of IAT, while expression of mono-nuclear cells (MNC) L-selectin slightly increased duringIAT. Expression of PMN Mac-1 was markedly increasedat the end of IAT, whereas expression of MNC Mac-1 didnot change during IAT. Leukocyte counts decreased 30min after starting IAT, and then increased to the initiallevel or higher in parallel with L-selectin downregulationand Mac-1 upregulation on PMN. L-selectin downregula-tion and Mac-1 upregulation on PMN suggested that acti-vation of PMN associated with changes in peripheral leu-kocyte counts occurred during IAT and might play somerole in modulating the human circulating blood andimmune systems. Key Words: Plasmapheresis—Immuno-adsorption therapy—Neuroimmunological disorder—L-selectin—Mac-1—Neutrophil.

In recent years, plasmapheresis has been estab-lished as a therapy for neuroimmunological disor-ders (1,2). Immunoadsorption therapy (IAT) usingan immunoadsorbent column which can adsorb andremove plasma proteins including immunoglobulins(3–5) has come to be used in the treatment of pa-tients with neuroimmunological disorders. IAT isthought to eliminate autoantibodies and immuno-complexes in plasma and is also suspected of modi-fying the circulating blood and immune systemsthrough mechanisms other than the elimination ofplasma proteins. Previously, we found and reportedthat transient decreases and subsequent increases inleukocyte counts were accompanied by induction ofseveral plasma chemotactic factors such as C3a, C5a,

beta-thromboglobulin (b-TG), platelet factor 4(PF-4), and interleukin 8 (IL-8) (6), which some-times might lead to respiratory failure if patientshad allergic reactions during IAT (7). However, afew papers that described the influence of IAT oncellular components in peripheral blood have beenreported.

Various kinds of adhesion molecules, selectin, in-tegrin, and immunoglobulin superfamilies are in-volved in leukocyte transmigration to inflammatorysites and leukocyte extravasation (8–12). Several in-vestigators have reported leukocyte activation withthe downregulation of leukocyte L-selectin(CD62L), one of the selectin family, and upregula-tion of leukocyte Mac-1 (CD11b/CD18), one of theintegrin family, during extracorporeal circulationsuch as cardiopulmonary bypass and hemodialysis(13–20). We therefore focused on changes inL-selectin and Mac-1 on leukocytes during IATand investigated whether leukocyte activation asso-ciated with changes in leukocyte counts occurs dur-ing IAT.

Received June 1999; revised January 2000Presented in part at the 2nd International Society for Apheresis

Congress, held April 15-18, 1999, in Saarbrucken, Germany.Address correspondence and reprint requests to Dr. Kohei Ota,

Department of Neurology, Neurological Institute, Tokyo Wom-en’s Medical University, Kawada-cho 8-1, Shinjuku-ku, Tokyo162-8666, Japan.

Therapeutic Apheresis4(3):229–234, Blackwell Science, Inc.© 2000 International Society for Apheresis

229

SUBJECTS AMD METHODS

Immunoadsorption therapySeveral sessions of IAT were conducted in 6 pa-

tients with neuroimmunological disorders (3 withchronic inflammatory demyelinating polyneuropa-thy [CIDP], 1 with multifocal motor neuropathy, 1with acute inflammatory demyelinating polyneurop-athy, and 1 with Sjogren syndrome with sensory neu-ropathy) using the Plasma Flow OP-05 (Asahi Medi-cal Corp., Tokyo, Japan) as a plasma separator andImmusorba TR-350 (Asahi Medical Corp., Tokyo,Japan) as an adsorption column. Two thousand to2,500 ml of plasma was treated in about 120 min ineach IAT session. Nafamostat mesilate (20–40 mg/h)was used as an anticoagulant. Heparinized periph-eral blood was drawn over time (30 min after, and oncompletion of IAT) to determine peripheral leuko-cyte counts and expressions of leukocyte CD62L andCD11b. Data sets for analysis in the study were ob-tained at the first IAT session of each patient.

Flow cytometryCD45-FITC and CD62L-PE or CD11b-PE

(Pharmingen, San Diego, CA, U.S.A.) were double-stained for 15 min at 4°C using whole peripheralblood to avoid mechanical stimulation by centrifu-gation, washing, and other procedures. Cytogramswere prepared based on side scattering and fluores-cence intensity of CD45-FITC. Mononuclear cells(MNC) and neutrophil (PMN) populations were se-lected on the cytograms obtained. The expression ofCD62L-PE or CD11b-PE in each population wasevaluated by EPICS Profile II (Coulter Corp., Hi-aleah, FL, U.S.A.).

Statistical analysisResults were expressed as mean ± SD, and the

continuous variables were analyzed by the Friedmantest.

RESULTS

No definite abnormal vital sign and symptom wasobserved in all the patients except for mild hypoten-sion during IAT. Sequential change of CD62L andCD11b expression on PMN and MNC during IAT ina patient with CIDP is shown in Fig. 1, representa-tively. Positive percentage and mean fluorescenceintensity (MFI) of PMN CD62L expression beforeIAT was 87.9% and 7.9 and decreased to 82.4% and6.9, respectively, at 30 min after starting IAT. A fur-ther decrease in CD62L expression on PMN was ob-served at 120 min after starting IAT (on completionof IAT). Positive percentage and MFI of CD62Lexpression on MNC slightly increased during IAT

(Fig. 1, left). Positive percentage and MFI of PMNCD11b expression before IAT was 83.0% and 6.1,and increased to 93.2% and 14.0, respectively, at 120min after starting IAT whereas no change of CDIIbexpression on MNC was observed during IAT (Fig.1, right).

Average CD62L expression on PMN in 6 patientswas positive in 73.3 ± 15.8% before IAT. This ratedecreased to 63.6 ± 29.6% at 30 min after startingIAT, then further decreased to 43.7 ± 17.1% at theend of IAT. Average MFI of CD62L expression onPMN also decreased to 7.2 ± 5.1 at 120 min afterstarting IAT from 11.9 ± 4.5 before IAT (Table 1).The sequential changes of CD62L expression, bothpositive percentage and MFI, on PMN during IATwere statistically significant (p 4 0.0155 and 0.0421,respectively). Average CD62L expression on MNCremained virtually unchanged but was slightly in-creased after IAT (Table 2). Average CD11b expres-sion on PMN in 6 patients increased with time duringIAT, and the positive expression rate significantly (p4 0.0302) increased to 93.9 ± 4.2% at the end of IATfrom 85.7 ± 14.9% before IAT (Table 1); however,average MFI of CD11b expression on PMN tendedto increase to 12.2 ± 8.3 at 120 min after starting IATform 5.0±1.3 before IAT (p 4 0.0695). The sequen-tial change of CD11b expression on MNC duringIAT showed no significance (Table 2).

Evaluation of blood samples obtained from theextracorporeal circulation system before and afterpassage through the plasma separator in a patientwith CIDP at 30 min after starting IAT showed thatthe positive expression rate of CD62L and CD11bon PMN were markedly changed from 82.4% and72.0% before passage to 21.4% and 92.2% after pas-sage, respectively.

Average peripheral leukocyte counts in 6 patientdecreased to 3,500 ± 2,200/mm3 at 30 min after start-ing IAT from the initial counts of 6,400 ± 2,100/mm3,and then significantly (p 4 0.0025) changed to13,000 ± 6,300/mm3 at 120 min after starting (at theend of IAT) in parallel with CD62L downregulationand CD11b upregulation on PMN (Table 3). Micro-scopic and flow cytometric investigations of leuko-cytes revealed that the majority of leukocytes thatchanged in their counts during IAT were PMN.

DISCUSSION

The first step of leukocyte migration from bloodvessels to tissues begins by contact between L-selectin molecules on the intravascular leukocytemembrane and selectin ligands on vascular endothe-lial cells (tethering/rolling). Leukocytes are activated

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Ther Apher, Vol. 4, No. 3, 2000

by this signal, resulting in increases in expression ofadhesion molecules such as Mac-1 on the leukocytemembrane (triggering). L-selectin is then rapidlyshed by the protease and disappears from the leuko-cyte membrane (selectin shedding), while activatedleukocytes strongly adhere to endothelial cells by theinteraction between adherent molecules includingMac-1 on leukocytes and their ligands such asICAM-1 on endothelial cells (adhesion). Finally, ac-tivated leukocytes penetrate and pass through theloose junction of vascular endothelial cells (transmi-gration). Thus, changes in certain adherent mol-ecules on leukocytes are closely related to the kinet-ics of leukocytes in blood vessels.

Investigators have reported leukocyte activationwith downregulation of PMN L-selectin andupregulation of PMN Mac-1 during extracor-poreal circulation such as cardiopulmonary by-pass and hemodialysis (13–20). Induction of comple-ments by the extracorporeal circulation systemwas thought to mainly cause leukocyte activationand subsequent peripheral leukocytopenia duringthese therapeutic conditions (21,22). Previously,our studies have documented increases in concentra-tion of not only activated complements C3a and C5a,but also b-TG, PF-4, and IL-8 associated with thetransient decreases and following increases inperipheral leukocyte counts through the course of

FIG. 1. Sequential change of CD62L (left) and CD11b (right) expressions was measured in neutrophils (top) and mononuclear cells(bottom) during IAT. The positive percentage and MFI of neutrophil CD62L expression decreased to 82.4% and 6.9 at 30 min after startingof IAT from 87.9% and 7.9 before IAT, respectively. A further decrease of neutrophil CD62L expression was observed at 120 min afterstarting of IAT or on completion of IAT. The positive percentage and MFI of mononuclear cell CD62L expression slightly increased duringIAT. On the other hand, the positive percentage and MFI of neutrophil CD11b expression increased to 93.2% and 14.0 at 120 min afterstarting of IAT or on completion of IAT from 83.0% and 6.1 before IAT, respectively. Mononuclear cell CD11b expression was notchanged during IAT. The histograms illustrate date from one experiment of a CIDP patient, representative of 6 such experiments. IAT:immunoadsorption therapy, Pcnt:percentage, MFI: mean fluorescence intensity, PMN: neutrophil, MNC: mononuclear cell.

NEUTROPHIL ACTIVATION 231


IAT session (6). L-selectin downregulation andMac-1 upregulation on PMN suggest PMN activa-tion therefore might be caused by a raised concen-tration of above chemotactic factors and probably bythe mechanical stimuli, for example, of high shearstress produced when they passed through theplasma separator.

More increases of peripheral leukocyte countsseemed to occur during IAT than those observedduring hemodialysis. The degree of PMN activationinduced by the IAT system, including an immuno-adsorption column and a plasma separator, might behigher than that by the hemodialysis system; how-ever, we observed PMN activation with low CD62Land high CD11b expression in one CIDP patientwhen blood has passed through the plasma separatorwithout the influence of the imunoadsorption col-umn. Further studies are needed to clarify the role of

the immunoadsorption column in activating leuko-cytes.

Nafamostat mesilate, expected to suppresscomplement activation, was used as an anticoagulantin the study. However, with our clinical dosage, it didnot prevent complement activation as well as withheparin sodium (6) nor PMN activation as statedabove. Because of the higher expression of adherentmolecules on PMN at the early stage of IAT, PMNmay first accumulate in the peripheral vessels includ-ing pulmonary capillaries and release various chemi-cal mediators quickly. IAT seems to reduce the peakexpiratory flow rate, a sensitive indicator for bron-chospasm, within 15 min of initiating the procedure,and may trigger an attack in patients with a historyof asthma (7), and leukocyte counts in peripheralblood would then decrease at that time. As the con-centrations of many chemotactic factors in the

TABLE 2. Positive percentage and mean fluorescence intensity of CD62L and CD11b on mononuclear cellsduring immunoadsorption therapy

Patient (age, sex) Diagnosis

CD62L % (MFI) CD11b % (MFI)

Before 30 min of IAT End of IAT Before 30 min of IAT End of IAT

KH (56, M) CIDP 20.5 (11.4) 20.4 (11.3) 19.5 (11.8) 24.2 (4.5) 18.1 (3.2) 37.3 (4.9)CM (59, F) CIDP 68.3 (11.1) 73.7 (11.0) 74.7 (11.8) 9.2 (2.6) 4.4 (2.3) 10.6 (2.8)HK (43, M) MMN 63.8 (12.1) 71.4 (13.0) 65.6 (15.0) 2.9 (2.7) 6.4 (2.6) 5.4 (3.1)KH (15, M) AIDP 51.7 (21.4) 52.8 (20.9) 49.1 (20.8) 9.7 (6.5) 9.3 (3.4) 10.3 (9.0)TH (51, M) CIDP 47.7 (18.7) 54.2 (19.1) 47.8 (17.1) 16.0 (6.3) 12.5 (5.4) 14.8 (4.6)KN (52, F) Sjs, neuropathy 62.9 (20.5) 68.8 (20.2) 60.2 (18.5) 5.7 (3.0) 11.0 (3.1) 17.6 (3.2)

Average 52.5 (15.9) 56.9 (15.9) 52.8a (15.8)c 11.3 (4.3) 10.3 (3.3) 16b (4.6)d

SD 17.5 (4.8) 20.0 (4.6) 19.2 (3.7) 7.7 (1.8) 4.8 (1.1) 11.2 (2.3)

The average percentage and mean fluorescence intensity of CD62L and CD11b on mononuclear cells showed no significance.a p 4 0.1353.b p 4 0.1146.c p 4 0.8465.d p 4 0.0695 (Friedman test).

TABLE 1. Positive percentage and mean fluorescence intensity of CD62L and CD11b on neutrophils during IAT


CD62L % (MFI) CD11b % (MFI)

Before 30 min of IAT End of IAT Before 30 min of IAT End of IAT

KH (56, M) CIDP 97.9 (10.2) 96.3 (12.8) 72.6 (2.5) 90.4 (5.4) 85.1 (4.5) 98.0 (22.4)CM (59, F) CIDP 87.9 (7.9) 82.4 (6.9) 39.3 (2.3) 83.0 (6.1) 72.0 (4.1) 93.2 (14.0)HK (43, M) MMN 59.8 (7.2) 10.9 (3.1) 31.9 (4.1) 56.8 (7.0) 83.7 (21.3) 86.6 (21.0)KH (15, M) AIDP 62.5 (14.0) 55.4 (16.5) 27.3 (12.7) 96.2 (3.8) 33.5 (3.4) 93.5 (5.3)TH (51, M) CIDP 68.7 (19.3) 74.0 (22.4) 55.3 (13.7) 93.6 (4.1) 97.1 (3.1) 97.8 (5.9)KN (52, F) Sjs, neuropathy 63.1 (12.8) 62.5 (12.6) 35.8 (7.9) 94.1 (3.8) 83.4 (3.0) 94.4 (3.8)

Average 73.3 (11.9) 63.6 (12.4) 43.7a (7.2)c 85.7 (5.0) 75.8 (6.6) 93.9b (12.2)d

SD 15.8 (4.5) 29.6 (6.8) 17.1 (5.1) 14.9 (1.3) 22.2 (7.2) 4.2 (8.3)

MFI: mean fluorescence intensity, IAT: immunoadsorption therapy, CIDP: chronic inflammatory demyelinating polyneuropathy, MMN:multifocal motor neuropathy, AIDP: acute inflammatory demyelinating polyneuropathy, Sjs: Sjogren syndrome, SD: standard deviation.

The average percentage and mean fluorescence intensity of CD62L on neutrophils of 6 patients statistically decreased during IAT byFriedman test. The average percentage of CD11b on neutrophils of 6 patients statistically increased during IAT by Freidman test.

a p 4 0.0155.b p 4 0.0302.c p 4 0.0421.d p 4 0.0695 (Friedman test).

K. OTA ET AL.232


plasma gradually increase during IAT and reach themaximum at 120 min or at the end of the IAT ses-sion, many PMN may be recruited again by a raisedconcentration of chemoattractants (23) in the blood-stream from the extravascular space, and leukocytecounts may return to the initial or higher level by 120min or around the end of IAT. After completion ofIAT, leukocyte counts become normalized immedi-ately due to mechanisms such as discontinuation ofleukocyte recruitment by chemotactic factors, andpossibly due to apotosis of activated PMN (24,25).

When we apply IAT for the treatment of neuro-immunological disorders, IAT seems to be effectivefor some patients of whom etiology is not thought tobe mediated by autoantibodies. Several mechanismsof IAT efficacy other than the elimination of immu-noglobulins and immune complexes are thereforespeculated. Recently PMN and lymphocyte interac-tion was also discussed, and activated PMN must beinfluenced by lymphocyte functions. Studies that de-fensin, released from IL-8–stimulated PMN, en-hanced T cell recruitment, and antigen-specific T cellresponse have been reported (26,27). Thus, such adramatic increase in activated PMN with changes inexpression of L-selectin and Mac-1 during IAT mayhave a potential of modulating cellular immunity.Further studies are needed to confirm the efficacy ofIAT for the human immune network.

CONCLUSIONS

L-selectin downregulation and Mac-1 upregulationon PMN suggested that PMN activation associatedwith changes in peripheral leukocyte counts oc-curred during IAT.

REFERENCES

1. The Guillain-Barre Syndrome Study Group. Plasmapheresisand acute Guillain-Barre syndrome. Neurology 1985;35:1096–104.

2. Dyck PJ, Daube J, O’Brien P, Pineda A, Low PA, Windebank

AJ, Swanson C. Plasma exchange in chronic inflammatorydemyelinating polyradiculoneuropathy. N Engl J Med 1986;314:461–5.

3. Yamazaki Z, Fujimori Y, Takahama T, Inoue N, Wada T,Kazama M, Morioka M, Abe T, Yamawaki N, Inagaki K.Efficiency and biocompatibility of a new immunosorbent.Trans Am Soc Artif Intern Organs 1982;28:318–23.

4. Shibuya N, Nagasato K, Shibuyama K, Kanazawa H. Immu-noadsorption therapy in neurologic disorders: Myastheniagravis, multiple sclerosis and Guillain-Barré-syndrome. In:Oda T, Shiokawa Y, Inoue N, eds. Therapeutic Plasmapher-esis (VI). Cleveland: ISAO Press, 1987:122–8

5. Heininger K, Toyka KV, Borberg H. Selective removal ofantibodies: Theoretical and practical aspect. In: Oda T, Shio-kawa Y, Inoue N, eds. Therapeutic Plasmapheresis (VI).Cleveland: ISAO Press, 1987:136–42

6. Ota K, Ikusaka M, Shimizu Y, Akiyama N, Iwata M. Increasein peripheral blood leukocytes and chemokines during immu-noadsorbent therapy. Jpn J Apheresis 1997;16:309–10.

7. Ikusaka M, Ota K, Ichikawa H, Ueda M, Iwata M. Transientdecrease in peak expiratory flow rate during immunoadsor-bent therapy. Jpn J Apheresis 1997;16:311–2.

8. Miller LJ, Schwarting R, Springer TA. Regulated expressionof the Mac-1, LFA-1, p150,95 glycoprotein family during leu-kocyte differentiation. J Immunol 1986;137:2891–900.

9. Springer TA. Adhesion receptors of the immune system. Na-ture 1990;346:425–34.

10. Larson RS, Springer TA. Structure and function of leukocyteintegrins. Immunol Rev 1990;114:181–217.

11. Picker LJ, Siegelman MH. Lymphoid tissues and organs. In:Paul WE, ed. Fundamental Immunology, third edition. NewYork: Raven Press Ltd, 1993:145–98.

12. Johnson PA, Alexander HD, McMillan SA, Maxwell AP. Up-regulation of the granulocyte adhesion molecule Mac-1 byautoantibodies in autoimmune vasculitis. Clin Exp Immunol1997;107:513–9.

13. Takala AJ, Jousela IT, Takkunen OS, Jansson SE, KyosolaKT, Olkkola KT, Leirisalo-Repo M, Repo H. Time course ofbeta 2-integrin CD11b/CD18 (Mac-1, alpha M, beta 2) up-regulation on neutrophils and monocytes after coronary ar-tery bypass grafting. CD11b upregulation after CABG sur-gery. Scand J Thorac Cardiovasc Surg 1996;30:141–8.

14. Himmelfarb J, Zaoui P, Hakim R. Modulation of granulocyteLAM-1 and MAC-1 during dialysis—A prospective, random-ized controlled trial. Kidney Int 1992;41:388–95.

15. Lundahl J, Hed J, Jacobson SH. Dialysis granulocytopenia ispreceded by an increased surface expression of the adhesion-promoting glycoprotein Mac-1. Nephron 1992;61:163–9.

16. Thylen P, Lundahl J, Fernvik E, Hed J, Svenson SB, JacobsonSH. Mobilization of an intracellular glycoprotein (Mac-1) onmonocytes and granulocytes during hemodialysis. Am JNephrol 1992;12:393–400.

17. Kappelmayer J, Bernabei A, Gikakis N, Edmunds LH Jr,

TABLE 3. Peripheral leukocyte counts during IAT


Leukocyte count/mm3 (PMN%)

Before 30 min of IAT End of IAT

KH (56, M) CIDP 8200 (87.2) 4600 (79.8) 21600 (78.6)CM (59, F) CIDP 4100 (n.d.) 2000 (n.d.) 5100 (n.d.)HK (43, M) MMN 7300 (n.d.) 4100 (n.d.) 19700 (n.d.)KH (15, M) AIDP 6200 (57.0) 2100 (18.5) 10600 (74.8)TH (51, M) CIDP 8700 (54.0) 7100 (49.7) 11300 (62.0)KN (52, F) Sjs, neuropathy 3600 (54.8) 1300 (27.0) 9800 (78.5)

Average 6400 3500 13000a

SD 2100 2200 6300

n.d.: not done.a p 4 0.0025 (Friedman test).

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Colman RW. Upregulation of Mac-1 surface expression onneutrophils during simulated extracorporeal circulation. JLab Clin Med 1993; 121:118–26.

18. Rabb H, Agosti SJ, Bittle PA, Fernandez M, Ramirez G,Tedder TF. Alterations in soluble and leukocyte surface L-selectin (CD62L) in hemodialysis patients. J Am Soc Nephrol1995;6:1445–50.

19. Kawabata K, Nagake Y, Shikata K, Makino H, Ota Z. Thechanges of Mac-1 and L-selectin expression on granulocytesand soluble L-selectin level during hemodialysis. Nephron1996;73:573–9.

20. Dou L, Brunet P, Dignat-George F, Sampol J, Berland Y.Effect of uremia and hemodialysis on soluble L-selectin andleukocyte surface CD11b and L-selectin. Am J Kidney Dis1998;31:67–73.

21. Craddock PR, Fehr J, Dalmasso AP, Brigham KL, Jacob HS.Hemodialysis leukopenia. Pulmonary vascular leukostasis re-sulting from complement activation by dialyzer cellophanemembranes. J Clin Invest 1977;59:879–88.

22. Shibuya N, Origuchi T. Plasmapheresis for autoimmune neu-rological disorder. Neurol Therap (Japan) 1990;7:397–403.

23. Durum SK, Oppenheim JJ. Proinflammatory cytokines andimmunity. In: Paul WE, ed. Fundamental Immunology, thirdedition. New York: Raven Press Ltd, 1993:801–35.

24. Watson RWG, Redmond HP, Wang JH, Condron C,Bouchier-Hayes D. Neutrophils undergo apoptosis followingingestion of Escherichia coli. J Immunol 1996;156:3986–92.

25. Meagher LC, Cousin JM, Seckl JR, Haslett C. Opposing ef-fects of glucocorticoids on the rate of apoptosis in neutro-philic and eosinophilic granulocytes. J Immunol 1996;156:4422–8.

26. Taub DD, Anver M, Oppenheim JJ, Longo DL, Murphy WJ.T lymphocyte recruitment by interleukin-8 (IL-8). IL-8-induced degranulation of neutrophils releases potent che-moattractants for human T lymphocytes both in vitro and invivo. J Clin Inv 1996;97:1931–41.

27. Chertov O, Michiel DF, Xu L, Wang JM, Tani K, Murphy WJ,Longo DL, Taub DD, Oppenheim JJ. Identification of defen-sin-1, defensin-2, and CAP37/azurocidin as T-cell chemoat-tractant proteins released from interleukin-8-stimulated neu-trophil. J Bio Chem 1996;271:2935–40.

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