Skip to main content
PMC home page
Annals of the Rheumatic Diseases logoLink to Annals of the Rheumatic Diseases
. 2000 Jul;59(7):555–564. doi: 10.1136/ard.59.7.555

Neutrophil function in pregnancy and rheumatoid arthritis

I Crocker 1, P Baker 1, J Fletcher 1
PMCID: PMC1753180  PMID: 10873967

Abstract

BACKGROUND—Pregnancy exerts suppressive effects on rheumatoid arthritis (RA). An attenuation in neutrophil function in late pregnancy which may explain this amelioration has previously been reported.
OBJECTIVE—A longitudinal investigation of neutrophil activity in healthy pregnant women (n=9) and pregnant patients with RA (n=9), compared with age matched non-pregnant patients with RA (n=12) and healthy controls (n=22).
METHODS—Neutrophil activation was measured in response to the physiological receptor agonists, n-formyl-methionyl-leucyl-phenylalanine (fMLP) and zymosan activated serum (ZAS). Superoxide anion production (respiratory burst) was determined by lucigenin enhanced chemiluminescence (LUCL); secondary granule lactoferrin release by enzyme linked immunosorbent assay (ELISA); and CD11b, CD18, and CD62L expression by flow cytometric analysis.
RESULTS—Stimulated neutrophil LUCL was significantly reduced in both pregnant women with RA and healthy pregnant women in the second (fMLP 43% and 69%, ZAS 43% and 59%, respectively) and third trimesters (fMLP 24% and 44%, ZAS 32% and 38%, respectively). Responses returned to normal within eight weeks of delivery and unstimulated levels remained unchanged throughout pregnancy. Basal and stimulated CD11b, CD18, and CD62L expression showed no variations throughout gestation for both pregnancy groups. Likewise, stimulated lactoferrin release and plasma lactoferrin remained unchanged. Certain morphological differences in RA neutrophils were highlighted by the flow cytometric analysis. Moreover, resting neutrophils and stimulated cells from patients with RA, including pregnant subjects, showed a marked increase in LUCL, but a reduction in CD11b, CD18, and CD62L. Low dose prednisolone and methylprednisolone had no effect on neutrophil parameters over the period of treatment with non-steroidal anti-inflammatory drugs.
CONCLUSION—The attenuation to neutrophil respiratory burst in both healthy and RA pregnancies may offer an explanation for the pregnancy induced remission of this inflammatory disorder.



Full Text

The Full Text of this article is available as a PDF (177.0 KB).

Figure 1  .

Figure 1  

Open in a new tab

Basal neutrophil lucigenin enhanced chemiluminescence and the effect of fMLP stimulation on cell responses from (A) healthy pregnant women (n=9) and non-pregnant controls matched for age (n=22); (B) pregnant RA women (n=9) and non-pregnant patients with RA matched for age (n=12). Integral chemiluminescence light output was recorded as relative light units (RLUs). The non-pregnant results are represent as shaded areas of means (SEM). Longitudinal raw data for pregnant subjects are represented by individual symbols. Stimulated cell responses above resting cell levels are indicated. During the second and third trimester of pregnancy, respiratory burst activity was significantly reduced for both pregnancy groups in response to fMLP and zymosan activated serum (ZAS) stimulation (ZAS results not shown). Neutrophil activity for both pregnancy groups returned to their respective control levels within seven weeks of delivery (see table 2).

Figure 2  .

Figure 2  

Open in a new tab

Expression of adhesion molecules on the surface of neutrophils from the four study groups. The results are given for CD11b, CD18, and CD62L and show changes in mean fluorescence units before stimulation (phosphate buffered saline) and after stimulation with fMLP or zymosan activated serum. Data are expressed as mean values (SEM). Statistical significance of difference was determined between the rheumatoid groups and their equivalent healthy, non-pregnant and pregnant control groups. The degree of statistical significance is indicated by *p<0.05 and **p<0.01 (t test).

Figure 3  .

Figure 3  

Open in a new tab

  Expression of CD62L on the surface of neutrophils from women of the four study groups. The results show changes in the percentage of positive cells present before stimulation (phosphate buffered saline) and after stimulation with fMLP or zymosan activated serum. Data are expressed as mean values (SEM).

Figure 4  .

Figure 4  

Open in a new tab

  Plasma lactoferrin and unstimulated (phosphate buffered saline) and stimulated (fMLP, zymosan activated serum) whole blood lactoferrin release from non-pregnant and pregnant healthy women, and from non-pregnant and pregnant patients with RA matched for age. Data are shown as mean (SEM) concentrations (nmol/l). Statistical significance of difference was determined between the RA groups and their equivalent healthy, non-pregnant and pregnant control groups. The degree of significance is indicated by *p<0.05 and **p<0.01 (t test).

Figure 5  .

Figure 5  

Open in a new tab

Lactoferrin concentrations normalised to 106 cells for plasma and unstimulated (phosphate buffered saline) and stimulated neutrophils (fMLP, zymosan activated serum) of healthy non-pregnant and pregnant women and RA, non-pregnant and pregnant women. The data are expressed as means (SEM) (pmol/106 cells).

Figure 6  .

Figure 6  

Open in a new tab

Morphological differences for neutrophils from healthy non-pregnant and pregnant women and RA non-pregnant and pregnant women as assessed by their light scattering properties on a flow cytometer. Forward scatter indicates cell size and side scatter shows cell complexity. The data are expressed as mean values (SEM). Statistical significance of difference was determined between the rheumatoid groups and their equivalent healthy, non-pregnant and pregnant control groups. The degree of significance is indicated by *p<0.05 and **p<0.01 (t test).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ANDRESEN R. H., MONROE C. W. Experimental study of the behavior of adult human skin homografts during pregnancy. A preliminary report. Am J Obstet Gynecol. 1962 Oct 15;84:1096–1103. doi: 10.1016/0002-9378(62)90560-4. [DOI] [PubMed] [Google Scholar]
  2. Adeyemi E. O., Campos L. B., Loizou S., Walport M. J., Hodgson H. J. Plasma lactoferrin and neutrophil elastase in rheumatoid arthritis and systemic lupus erythematosus. Br J Rheumatol. 1990 Feb;29(1):15–20. doi: 10.1093/rheumatology/29.1.15. [DOI] [PubMed] [Google Scholar]
  3. Baggiolini M., Walz A., Kunkel S. L. Neutrophil-activating peptide-1/interleukin 8, a novel cytokine that activates neutrophils. J Clin Invest. 1989 Oct;84(4):1045–1049. doi: 10.1172/JCI114265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bainton D. F., Miller L. J., Kishimoto T. K., Springer T. A. Leukocyte adhesion receptors are stored in peroxidase-negative granules of human neutrophils. J Exp Med. 1987 Dec 1;166(6):1641–1653. doi: 10.1084/jem.166.6.1641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barden A., Graham D., Beilin L. J., Ritchie J., Baker R., Walters B. N., Michael C. A. Neutrophil CD11B expression and neutrophil activation in pre-eclampsia. Clin Sci (Lond) 1997 Jan;92(1):37–44. doi: 10.1042/cs0920037. [DOI] [PubMed] [Google Scholar]
  6. Baynes R. D., Bezwoda W. R., Khan Q., Mansoor N. Relationship of plasma lactoferrin content to neutrophil regeneration and bone marrow infusion. Scand J Haematol. 1986 Jan;36(1):79–84. doi: 10.1111/j.1600-0609.1986.tb02654.x. [DOI] [PubMed] [Google Scholar]
  7. Berkow R. L., Wang D., Larrick J. W., Dodson R. W., Howard T. H. Enhancement of neutrophil superoxide production by preincubation with recombinant human tumor necrosis factor. J Immunol. 1987 Dec 1;139(11):3783–3791. [PubMed] [Google Scholar]
  8. Björksten B., Söderström T., Damber M. G., von Schoultz B., Stigbrand T. Polymorphonuclear leucocyte function during pregnancy. Scand J Immunol. 1978;8(3):257–262. doi: 10.1111/j.1365-3083.1978.tb00518.x. [DOI] [PubMed] [Google Scholar]
  9. Bond A., Hay F. C. L-selectin expression on the surface of peripheral blood leucocytes from rheumatoid arthritis patients is linked to disease activity. Scand J Immunol. 1997 Sep;46(3):312–316. doi: 10.1046/j.1365-3083.1997.d01-131.x. [DOI] [PubMed] [Google Scholar]
  10. Brabin B. J. Epidemiology of infection in pregnancy. Rev Infect Dis. 1985 Sep-Oct;7(5):579–603. doi: 10.1093/clinids/7.5.579. [DOI] [PubMed] [Google Scholar]
  11. Brown R. D., Yuen E., Rickard K. A., Vincent P. C., Young G., Kronenberg H. Plasma lactoferrin in patients with neutropenia. Blut. 1986 May;52(5):289–295. doi: 10.1007/BF00320792. [DOI] [PubMed] [Google Scholar]
  12. Buyon J. P., Korchak H. M., Rutherford L. E., Ganguly M., Weissmann G. Female hormones reduce neutrophil responsiveness in vitro. Arthritis Rheum. 1984 Jun;27(6):623–630. doi: 10.1002/art.1780270604. [DOI] [PubMed] [Google Scholar]
  13. Cotton D. J., Seligmann B., O'Brien W. F., Gallin J. I. Selective defect in human neutrophil superoxide anion generation elicited by the chemoattractant N-formylmethionylleucylphenylalanine in pregnancy. J Infect Dis. 1983 Aug;148(2):194–199. doi: 10.1093/infdis/148.2.194. [DOI] [PubMed] [Google Scholar]
  14. Crouch S. P., Crocker I. P., Fletcher J. The effect of pregnancy on polymorphonuclear leukocyte function. J Immunol. 1995 Dec 1;155(11):5436–5443. [PubMed] [Google Scholar]
  15. Crouch S. P., Fletcher J. Effect of ingested pentoxifylline on neutrophil superoxide anion production. Infect Immun. 1992 Nov;60(11):4504–4509. doi: 10.1128/iai.60.11.4504-4509.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Da Silva J. A., Spector T. D. The role of pregnancy in the course and aetiology of rheumatoid arthritis. Clin Rheumatol. 1992 Jun;11(2):189–194. doi: 10.1007/BF02207955. [DOI] [PubMed] [Google Scholar]
  17. Edwards S. W., Hughes V., Barlow J., Bucknall R. Immunological detection of myeloperoxidase in synovial fluid from patients with rheumatoid arthritis. Biochem J. 1988 Feb 15;250(1):81–85. doi: 10.1042/bj2500081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. El-Maallem H., Fletcher J. Impaired neutrophil function and myeloperoxidase deficiency in pregnancy. Br J Haematol. 1980 Mar;44(3):375–381. doi: 10.1111/j.1365-2141.1980.tb05906.x. [DOI] [PubMed] [Google Scholar]
  19. Emery P., Lopez A. F., Burns G. F., Vadas M. A. Synovial fluid neutrophils of patients with rheumatoid arthritis have membrane antigen changes that reflect activation. Ann Rheum Dis. 1988 Jan;47(1):34–39. doi: 10.1136/ard.47.1.34. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Ertuğ H., Arman M., Yeğin O. Neutrophil, monocyte and lymphocyte locomotion in rheumatic fever and rheumatoid arthritis. Turk J Pediatr. 1990 Apr-Jun;32(2):73–78. [PubMed] [Google Scholar]
  21. Esaguy N., Aguas A. P., Vilanova M., Silva M. T. Activation of human neutrophils by phorbol ester decreases the cytoplasm compactness and the lactoferrin content of the granulocytes. J Leukoc Biol. 1991 Nov;50(5):444–452. doi: 10.1002/jlb.50.5.444. [DOI] [PubMed] [Google Scholar]
  22. Espersen G. T., Ernst E., Vestergaard M., Pedersen J. O., Grunnet N. Changes in PMN leukocyte migration activity and complement C3d levels in RA patients with high disease activity during steroid treatment. Scand J Rheumatol. 1989;18(1):51–56. doi: 10.3109/03009748909095403. [DOI] [PubMed] [Google Scholar]
  23. Felzmann T., Gadd S., Majdic O., Maurer D., Petera P., Smolen J., Knapp W. Analysis of function-associated receptor molecules on peripheral blood and synovial fluid granulocytes from patients with rheumatoid and reactive arthritis. J Clin Immunol. 1991 Jul;11(4):205–212. doi: 10.1007/BF00917426. [DOI] [PubMed] [Google Scholar]
  24. Fernandez H. N., Henson P. M., Otani A., Hugli T. E. Chemotactic response to human C3a and C5a anaphylatoxins. I. Evaluation of C3a and C5a leukotaxis in vitro and under stimulated in vivo conditions. J Immunol. 1978 Jan;120(1):109–115. [PubMed] [Google Scholar]
  25. Finn R., St Hill C. A., Govan A. J., Ralfs I. G., Gurney F. J., Denye V. Immunological responses in pregnancy and survival of fetal homograft. Br Med J. 1972 Jul 15;3(5819):150–152. doi: 10.1136/bmj.3.5819.150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Gyllenhammar H. Lucigenin chemiluminescence in the assessment of neutrophil superoxide production. J Immunol Methods. 1987 Mar 12;97(2):209–213. doi: 10.1016/0022-1759(87)90461-3. [DOI] [PubMed] [Google Scholar]
  27. Jones J., Laffafian I., Cooper A. M., Williams B. D., Morgan B. P. Expression of complement regulatory molecules and other surface markers on neutrophils from synovial fluid and blood of patients with rheumatoid arthritis. Br J Rheumatol. 1994 Aug;33(8):707–712. doi: 10.1093/rheumatology/33.8.707. [DOI] [PubMed] [Google Scholar]
  28. Kleesiek K., Reinards R., Brackertz D., Neumann S., Lang H., Greiling H. Granulocyte elastase as a new biochemical marker in the diagnosis of chronic joint diseases. Rheumatol Int. 1986;6(4):161–169. doi: 10.1007/BF00541283. [DOI] [PubMed] [Google Scholar]
  29. Krause P. J., Ingardia C. J., Pontius L. T., Malech H. L., LoBello T. M., Maderazo E. G. Host defense during pregnancy: neutrophil chemotaxis and adherence. Am J Obstet Gynecol. 1987 Aug;157(2):274–280. doi: 10.1016/s0002-9378(87)80150-3. [DOI] [PubMed] [Google Scholar]
  30. Kremer J. M. Possible mechanisms of action of methotrexate in patients with rheumatoid arthritis. Br J Rheumatol. 1995 Nov;34 (Suppl 2):26–29. [PubMed] [Google Scholar]
  31. Macey M. G., Wilton J. M., Carbon R., Edmonds S., Perry J. D., McCarthy D. Leukocyte activation and function-associated antigens in inflammatory disease. Agents Actions. 1993;38(Spec No):C39–C40. doi: 10.1007/BF01991130. [DOI] [PubMed] [Google Scholar]
  32. Malmquist J., Thorell J. I., Wolheim F. A. Lactoferrin and lysozyme in arthritic exudates. Acta Med Scand. 1977;202(4):313–318. doi: 10.1111/j.0954-6820.1977.tb16834.x. [DOI] [PubMed] [Google Scholar]
  33. Manicourt D. H., Triki R., Fukuda K., Devogelaer J. P., Nagant de Deuxchaisnes C., Thonar E. J. Levels of circulating tumor necrosis factor alpha and interleukin-6 in patients with rheumatoid arthritis. Relationship to serum levels of hyaluronan and antigenic keratan sulfate. Arthritis Rheum. 1993 Apr;36(4):490–499. doi: 10.1002/art.1780360409. [DOI] [PubMed] [Google Scholar]
  34. Merry P., Winyard P. G., Morris C. J., Grootveld M., Blake D. R. Oxygen free radicals, inflammation, and synovitis: and synovitis: the current status. Ann Rheum Dis. 1989 Oct;48(10):864–870. doi: 10.1136/ard.48.10.864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Miller C., Russell A. S. Polymorphonuclear leukocyte response to stimulation in vitro during pregnancy. Inflammation. 1986 Sep;10(3):215–222. doi: 10.1007/BF00916117. [DOI] [PubMed] [Google Scholar]
  36. Mur E., Zabernigg A., Hilbe W., Eisterer W., Halder W., Thaler J. Oxidative burst of neutrophils in patients with rheumatoid arthritis: influence of various cytokines and medication. Clin Exp Rheumatol. 1997 May-Jun;15(3):233–237. [PubMed] [Google Scholar]
  37. Nelson J. L., Hughes K. A., Smith A. G., Nisperos B. B., Branchaud A. M., Hansen J. A. Maternal-fetal disparity in HLA class II alloantigens and the pregnancy-induced amelioration of rheumatoid arthritis. N Engl J Med. 1993 Aug 12;329(7):466–471. doi: 10.1056/NEJM199308123290704. [DOI] [PubMed] [Google Scholar]
  38. Nurcombe H. L., Bucknall R. C., Edwards S. W. Neutrophils isolated from the synovial fluid of patients with rheumatoid arthritis: priming and activation in vivo. Ann Rheum Dis. 1991 Mar;50(3):147–153. doi: 10.1136/ard.50.3.147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Persellin R. H. The effect of pregnancy on rheumatoid arthritis. Bull Rheum Dis. 1976;27(9):922–927. [PubMed] [Google Scholar]
  40. Persellin R. H., Thoi L. L. Human polymorphonuclear leukocyte phagocytosis in pregnancy. Development of inhibition during gestation and recovery in the postpartum period. Am J Obstet Gynecol. 1979 Jun 1;134(3):250–255. doi: 10.1016/s0002-9378(16)33028-9. [DOI] [PubMed] [Google Scholar]
  41. Selvaraj R. J., Sbarra A. J., Thomas G. B., Cetrulo C. L., Mitchell G. W., Jr A microtechnique for studying chemiluminescence response of phagocytes using whole blood and its application to the evaluation of phagocytes in pregnancy. J Reticuloendothel Soc. 1982 Jan;31(1):3–16. [PubMed] [Google Scholar]
  42. Spector T. D., Da Silva J. A. Pregnancy and rheumatoid arthritis: an overview. Am J Reprod Immunol. 1992 Oct-Dec;28(3-4):222–225. doi: 10.1111/j.1600-0897.1992.tb00797.x. [DOI] [PubMed] [Google Scholar]
  43. Spertini O., Kansas G. S., Munro J. M., Griffin J. D., Tedder T. F. Regulation of leukocyte migration by activation of the leukocyte adhesion molecule-1 (LAM-1) selectin. Nature. 1991 Feb 21;349(6311):691–694. doi: 10.1038/349691a0. [DOI] [PubMed] [Google Scholar]
  44. Stanworth D. R. A possible immunochemical explanation for pregnancy associated remissions in rheumatoid arthritis? Ann Rheum Dis. 1988 Feb;47(2):89–90. doi: 10.1136/ard.47.2.89. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Storgaard M., Jensen M. P., Stengaard-Pedersen K., Møller B. K., Anderson P. L., Obel N. Effects of methotrexate, sulphasalazine and aurothiomalate on polymorphonuclear leucocytes in rheumatoid arthritis. Scand J Rheumatol. 1996;25(3):168–173. doi: 10.3109/03009749609080009. [DOI] [PubMed] [Google Scholar]
  46. Turzanski J., Crouch S. P., Fletcher J., Hunter A. Ex vivo neutrophil function in response to three different doses of glycosylated rHuG-CSF (Lenograstim). Br J Haematol. 1997 Jan;96(1):46–54. doi: 10.1046/j.1365-2141.1997.d01-2000.x. [DOI] [PubMed] [Google Scholar]
  47. Youssef P. P., Cormack J., Evill C. A., Peter D. T., Roberts-Thomson P. J., Ahern M. J., Smith M. D. Neutrophil trafficking into inflamed joints in patients with rheumatoid arthritis, and the effects of methylprednisolone. Arthritis Rheum. 1996 Feb;39(2):216–225. doi: 10.1002/art.1780390207. [DOI] [PubMed] [Google Scholar]
  48. Youssef P., Roberts-Thomson P., Ahern M., Smith M. Pulse methylprednisolone in rheumatoid arthritis: effects on peripheral blood and synovial fluid neutrophil surface phenotype. J Rheumatol. 1995 Nov;22(11):2065–2071. [PubMed] [Google Scholar]
  49. Zangerle P. F., De Groote D., Lopez M., Meuleman R. J., Vrindts Y., Fauchet F., Dehart I., Jadoul M., Radoux D., Franchimont P. Direct stimulation of cytokines (IL-1 beta, TNF-alpha, IL-6, IL-2, IFN-gamma and GM-CSF) in whole blood: II. Application to rheumatoid arthritis and osteoarthritis. Cytokine. 1992 Nov;4(6):568–575. doi: 10.1016/1043-4666(92)90021-i. [DOI] [PubMed] [Google Scholar]

Articles from Annals of the Rheumatic Diseases are provided here courtesy of BMJ Publishing Group

RESOURCES