Discontinuous density gradient analysis of human bone marrow: Presence of alloantigen-responsive, PHA-unresponsive cells in normal bone marrow; Absence of B lymphocytes in the bone marrow of patients with X-linked agammaglobulinemia

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Abstract

Normal human bone marrow cells were separated on discontinuous density gradients of 17–35% bovine serum albumin (BSA) into nine fractions. The top portion of the gradient (fractions 1–3, 17–23% BSA) was rich in cells characterized by a high rate of DNA synthesis and by the ability to give rise to colony-forming units. The lower portion of the gradient (fractions 5–9, 27–35% BSA) was rich in B lymphocytes capable of staining with a fluorescent antiserum to anti-human μ-chain, of forming rosettes with EAC1423, of taking up antigen, and of proliferating in response to pokeweed mitogen (PWM). Cells in the middle portion of the gradient (fractions 3–5, 21–27% BSA) showed the strongest reactivity in response to alloantigens but only an occasional weak reactivity to phytohemagglutinin (PHA). No E rosette-forming cells were found in any of the bone marrow fractions.

Bone marrow from two boys with proved X-linked agammaglobulinemia (X-LA) and one boy with probable X-LA was found to be totally deficient in B lymphocytes, as it lacked completely the cells identified by immunofluorescent staining and rosette formation with EAC1423, and failed to transform with PWM. Like normal marrow, marrow from patients with X-LA contained cells capable of proliferating in response to alloantigens but not in response to PHA.

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    Present address: Immunology Division, Department of Pediatrics, American University, Beirut, Lebanon.

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