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. 1973 Dec;52(12):3180–3189. doi: 10.1172/JCI107518

Differentiation Capacity of Cultured B Lymphocytes from Immunodeficient Patients

L Y F Wu 1,2, A R Lawton 1,2, M D Cooper 1,2
PMCID: PMC302594  PMID: 4543023

Abstract

Peripheral blood lymphocytes from 27 healthy individuals and from 18 patients with a diverse spectrum of defects in humoral immunity were examined for their capacity to undergo terminal differentiation in vitro. Pokeweed mitogen induced cells from normal persons to synthesize and secrete IgM. IgG, and IgA as detected by Immunofluorescence and incorporation of [14C]amino acids, Lymphocytes from three boys with X-linked agammaglobulinemia were stimulated to proliferate, but did not synthesize immunoglobulin. Lymphocyte cultures from three of four patients having agammaglobulinemia with B lymphocytes produced different immunoglobulin classes in ratios similar to the in vivo distribution of classes of B lymphocytes, Lymphocytes from a dysgammaglobulinemic boy deficient in serum IgG and IgA, but who had normal numbers of IgM-, IgG-, and IgA-bearing B lymphocytes, could not be stimulated by pokeweed mitogen to make IgG and IgA. Synthesis and secretion of IgA, as well as IgM and IgG, was detected in cell cultures from each of 10 patients with isolated IgA deficiency. The results suggest that deficiencies in immunoglobulin synthesis may reflect either (a) failure to develop B lymphocytes, (b) arrested development of B lymphocytes due to intrinsic metabolic abnormalities, or (c) disturbance of factors extrinsic to the B lymphocyte which are essential for normal induction of plasma cell maturation.

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Selected References

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  1. ASKONAS B. A., HUMPHREY J. H. Formation of antibody by isolated perfused lungs of immunized rabbits: the use of [14C]amino acids to study the dynamics of antibody secretion. Biochem J. 1958 Oct;70(2):212–222. doi: 10.1042/bj0700212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aiuti F., Fontana L., Gatti R. A. Membrane-bound immunoglobulin (Ig) and in vitro production of Ig by lymphoid cells from patients with primary immunodeficiencies. Scand J Immunol. 1973;2(1):9–16. doi: 10.1111/j.1365-3083.1973.tb02010.x. [DOI] [PubMed] [Google Scholar]
  3. Bankhurst A. D., Warner N. L. Surface immunoglobulins on the thoracic duct lymphocytes of the congenitally athymic (nude) mouse. Aust J Exp Biol Med Sci. 1972 Oct;50(5):661–664. doi: 10.1038/icb.1972.58. [DOI] [PubMed] [Google Scholar]
  4. Barker B. E., Farnes P., LaMarche P. H. Peripheral blood plasmacytosis following systemic exposure to Phytolacca americana (pokweed). Pediatrics. 1966 Sep;38(3):490–493. [PubMed] [Google Scholar]
  5. Byrt P., Ada G. L. An in vitro reaction between labelled flagellin or haemocyanin and lymphocyte-like cells from normal animals. Immunology. 1969 Oct;17(4):503–516. [PMC free article] [PubMed] [Google Scholar]
  6. COONS A. H., LEDUC E. H., CONNOLLY J. M. Studies on antibody production. I. A method for the histochemical demonstration of specific antibody and its application to a study of the hyperimmune rabbit. J Exp Med. 1955 Jul 1;102(1):49–60. doi: 10.1084/jem.102.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chessin L. N., Börjeson J., Welsh P. D., Douglas S. D., Cooper H. L. Studies on human peripheral blood lymphocytes in vitro. II. Morphological and biochemical studies on the transformation of lymphocytes by pokeweed mitogen. J Exp Med. 1966 Nov 1;124(5):873–884. doi: 10.1084/jem.124.5.873. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Choi Y. S., Biggar W. D., Good R. A. Biosynthesis and secretion of immunoglobulins by peripheral-blood lymphocytes in severe hypogammaglobulinaemia. Lancet. 1972 May 27;1(7761):1149–1152. doi: 10.1016/s0140-6736(72)91374-8. [DOI] [PubMed] [Google Scholar]
  9. Coombs R. R., Feinstein A., Wilson A. B. Immunoglobulin determinants on the surface of human lymphocytes. Lancet. 1969 Nov 29;2(7631):1157–1160. doi: 10.1016/s0140-6736(69)92484-2. [DOI] [PubMed] [Google Scholar]
  10. Cooper M. D., Lawton A. R., Bockman D. E. Agammaglobulinaemia with B lymphocytes. Specific defect of plasma-cell differentiation. Lancet. 1971 Oct 9;2(7728):791–794. doi: 10.1016/s0140-6736(71)92742-5. [DOI] [PubMed] [Google Scholar]
  11. Cooper M. D., Lawton A. R. Circulating B-cells in patients with immunodeficiency. Am J Pathol. 1972 Dec;69(3):513–528. [PMC free article] [PubMed] [Google Scholar]
  12. Cooper M. D., Lawton A. R., Kincade P. W. A two-stage model for development of antibody-producing cells. Clin Exp Immunol. 1972 May;11(1):143–149. [PMC free article] [PubMed] [Google Scholar]
  13. Crewther P., Warner N. L. Serum immunoglobulins and antibodies in congenitally athymic (nude) mice. Aust J Exp Biol Med Sci. 1972 Oct;50(5):625–635. doi: 10.1038/icb.1972.55. [DOI] [PubMed] [Google Scholar]
  14. Douglas S. D., Fudenberg H. H. In vitro development of plasma cells from lymphocytes following pokeweed mitogen stimulation: a fine structural study. Exp Cell Res. 1969 Feb;54(2):277–279. doi: 10.1016/0014-4827(69)90252-3. [DOI] [PubMed] [Google Scholar]
  15. Douglas S. D., Hoffman P. F., Borjeson J., Chessin L. N. Studies on human peripheral blood lymphocytes in vitro. 3. Fine structural features of lymphocyte transformation by pokeweed mitogen. J Immunol. 1967 Jan;98(1):17–30. [PubMed] [Google Scholar]
  16. Dwyer J. M., Hosking C. S. Antigen-binding lymphocytes in children with immune deficiency. Clin Exp Immunol. 1972 Oct;12(2):161–169. [PMC free article] [PubMed] [Google Scholar]
  17. Fröland S., Natvig J. B., Berdal P. Surface-bound immunoglobulin as a marker of B lymphocytes in man. Nat New Biol. 1971 Dec 22;234(51):251–252. doi: 10.1038/newbio234251a0. [DOI] [PubMed] [Google Scholar]
  18. Gajl-Peczalska K. J., Park B. H., Biggar W. D., Good R. A. B and T lymphocytes in primary immunodeficiency disease in man. J Clin Invest. 1973 Apr;52(4):919–928. doi: 10.1172/JCI107257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Geha R. S., Rosen F. S., Merler E. Identification and characterization of subpopulations of lymphocytes in human peripheral blood after fractionation on discontinuous gradients of albumin. The cellular defect in X-linked agammaglobulinemia. J Clin Invest. 1973 Jul;52(7):1726–1734. doi: 10.1172/JCI107354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Greaves M. F., Bauminger S., Janossy G. Lymphocyte activation. 3. Binding sites for phytomitogens on lymphocyte subpopulations. Clin Exp Immunol. 1972 Mar;10(3):537–554. [PMC free article] [PubMed] [Google Scholar]
  21. Greaves M. F. Biological effects of anti-immunoglobulins: evidence for immunoglobulin receptors on 'T' and 'B' lymphocytes. Transplant Rev. 1970;5:45–75. doi: 10.1111/j.1600-065x.1970.tb00356.x. [DOI] [PubMed] [Google Scholar]
  22. Greaves M. F., Roitt I. M. The effect of phytohaemagglutinin and other lymphocyte mitogens on immunoglobulin synthesis by human peripheral blood lymphocytes in vitro. Clin Exp Immunol. 1968 Jun;3(5):393–412. [PMC free article] [PubMed] [Google Scholar]
  23. Grey H. M., Rabellino E., Pirofsky B. Immunoglobulins on the surface of lymphocytes. IV. Distribution in hypogammaglobulinemia, cellular immune deficiency, and chronic lymphatic leukemia. J Clin Invest. 1971 Nov;50(11):2368–2375. doi: 10.1172/JCI106735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. HELMREICH E., KERN M., EISEN H. N. Observations on the mechanism of secretion of gamma-globulins by isolated lymph node cells. J Biol Chem. 1962 Jun;237:1925–1931. [PubMed] [Google Scholar]
  25. HELMREICH E., KERN M., EISEN H. N. The secretion of antibody by isolated lymph node cells. J Biol Chem. 1961 Feb;236:464–473. [PubMed] [Google Scholar]
  26. Hartzman R. J., Bach M. L., Bach F. H., Thurman G. B., Sell K. W. Precipitation of radioactively labeled samples: a semi-automatic multiple-sample processor. Cell Immunol. 1972 Jun;4(2):182–186. doi: 10.1016/0008-8749(72)90018-4. [DOI] [PubMed] [Google Scholar]
  27. Hijmans W., Schuit H. R., Klein F. An immunofluorescence procedure for the detection of intracellular immunoglobulins. Clin Exp Immunol. 1969 Apr;4(4):457–472. [PMC free article] [PubMed] [Google Scholar]
  28. Janossy G., Greaves M. F. Lymphocyte activation. I. Response of T and B lymphocytes to phytomitogens. Clin Exp Immunol. 1971 Oct;9(4):483–498. [PMC free article] [PubMed] [Google Scholar]
  29. Kincade P. W., Lawton A. R., Cooper M. D. Restriction of surface immunoglobulin determinants to lymphocytes of the plasma cell line. J Immunol. 1971 May;106(5):1421–1423. [PubMed] [Google Scholar]
  30. Lawton A. R., Royal S. A., Self K. S., Cooper M. D. IgA determinants on B-lymphocytes in patients with deficiency of circulating IgA. J Lab Clin Med. 1972 Jul;80(1):26–33. [PubMed] [Google Scholar]
  31. Marchalonis J. J., Cone R. E., Atwell J. L. Isolation and partial characterization of lymphocyte surface immunoglobulins. J Exp Med. 1972 Apr 1;135(4):956–971. doi: 10.1084/jem.135.4.956. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. McConnell I., Munro A., Gurner B. W., Coombs R. R. Studies on actively allergized cells. I. The cyto-dynamics and morphology of rosete-forming lymph node cells in mice and inhibition of rosette-formation with antibody to mouse immunoglobulins. Int Arch Allergy Appl Immunol. 1969;35(3):209–227. [PubMed] [Google Scholar]
  33. Oppenheim J. J., Rogentine G. N., Terry W. D. The transformation of human lymphocytes by monkey antisera to human immunoglobulins. Immunology. 1969 Jan;16(1):123–138. [PMC free article] [PubMed] [Google Scholar]
  34. Parkhouse R. M., Janossy G., Greaves M. F. Selective stimulation of IgM synthesis in mouse B lymphocytes by pokeweed mitogen. Nat New Biol. 1972 Jan 5;235(53):21–23. doi: 10.1038/newbio235021a0. [DOI] [PubMed] [Google Scholar]
  35. Pernis B., Forni L., Amante L. Immunoglobulin spots on the surface of rabbit lymphocytes. J Exp Med. 1970 Nov;132(5):1001–1018. doi: 10.1084/jem.132.5.1001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Preud'Homme J. L., Griscelli C., Seligmann M. Immunoglobulins on the surface of lymphocytes in fifty patients with primary immunodeficiency diseases. Clin Immunol Immunopathol. 1973 Jan;1(2):241–256. doi: 10.1016/0090-1229(73)90025-1. [DOI] [PubMed] [Google Scholar]
  37. Preud'homme J. L., Seligmann M. Primary immunodeficiency with increased numbers of circulating B lymphocytes contrasting with hypogammaglobulinaemia. Lancet. 1972 Feb 19;1(7747):442–442. doi: 10.1016/s0140-6736(72)90898-7. [DOI] [PubMed] [Google Scholar]
  38. Pritchard H., Riddaway J., Micklem H. S. Immune responses in congenitally thymus-less mice. II. Quantitative studies of serum immunoglobulins, the antibody response to sheep erythrocytes, and the effect of thymus allografting. Clin Exp Immunol. 1973 Jan;13(1):125–138. [PMC free article] [PubMed] [Google Scholar]
  39. Rabellino E., Colon S., Grey H. M., Unanue E. R. Immunoglobulins on the surface of lymphocytes. I. Distribution and quantitation. J Exp Med. 1971 Jan 1;133(1):156–167. doi: 10.1084/jem.133.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Raff M. C. Two distinct populations of peripheral lymphocytes in mice distinguishable by immunofluorescence. Immunology. 1970 Oct;19(4):637–650. [PMC free article] [PubMed] [Google Scholar]
  41. Roelants G. E., Askonas B. A. Immunological B memory in thymus deprived mice. Nat New Biol. 1972 Sep 13;239(89):63–64. doi: 10.1038/newbio239063a0. [DOI] [PubMed] [Google Scholar]
  42. Rowe D. S., Hug K., Faulk W. P., McCormick J. N., Gerber H. IgD on the surface of peripheral blood lymphocytes of the human newborn. Nat New Biol. 1973 Apr 4;242(118):155–157. doi: 10.1038/newbio242155a0. [DOI] [PubMed] [Google Scholar]
  43. SELL S., GELL P. G. STUDIES ON RABBIT LYMPHOCYTES IN VITRO. I. STIMULATION OF BLAST TRANSFORMATION WITH AN ANTIALLOTYPE SERUM. J Exp Med. 1965 Aug 1;122:423–440. doi: 10.1084/jem.122.2.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Stites D. P., Levin A. S., Austin K. E., Fudenberg H. H. Immunobiology of human lymphoid cell lines. I. Immunoglobulin biosynthesis in cultures from Hypogammaglobulinemias and paraproteinemias. J Immunol. 1971 Nov;107(5):1376–1381. [PubMed] [Google Scholar]
  45. Stockman G. D., Gallagher M. T., Heim L. R., South M. A., Trentin J. J. Differential stimulation of mouse lymphoid cells by phytohemagglutinin and pokeweed mitogen. Proc Soc Exp Biol Med. 1971 Mar;136(3):980–982. doi: 10.3181/00379727-136-35410. [DOI] [PubMed] [Google Scholar]
  46. Uhr J. W., Vitetta E. S. Synthesis, biochemistry and dynamics of cell surface immunoglobulin on lymphocytes. Fed Proc. 1973 Jan;32(1):35–40. [PubMed] [Google Scholar]
  47. Wilson J. D., Nossal G. J., Lewis H. Metabolic characteristics of lymphocyte surface immunoglobulins. Eur J Immunol. 1972 Jun;2(3):225–232. doi: 10.1002/eji.1830020306. [DOI] [PubMed] [Google Scholar]

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