Neonatal bone marrow transplantation of ADA-deficient SCID mice results in immunologic reconstitution despite low levels of engraftment and an absence of selective donor T lymphoid expansion

Adenosine deaminase (ADA)–deficient severe combined immune deficiency (SCID) may be treated by allogeneic hematopoietic stem cell transplantation without prior cytoreductive conditioning, although the mechanism of immune reconstitution is unclear. We studied this process in a murine gene knockout model of ADA-deficient SCID. Newborn ADA-deficient pups received transplants of intravenous infusion of normal congenic bone marrow, without prior cytoreductive conditioning, which resulted in long-term survival, multisystem correction, and nearly normal lymphocyte numbers and mitogenic proliferative responses. Only 1% to 3% of lymphocytes and myeloid cells were of donor origin without a selective expansion of donor-derived lymphocytes; immune reconstitution was by endogenous, host-derived ADA-deficient lymphocytes. Preconditioning of neonates with 100 to 400 cGy of total body irradiation before normal donor marrow transplant increased the levels of engrafted donor cells in a radiation dose–dependent manner, but the chimerism levels were similar for lymphoid and myeloid cells. The absence of selective reconstitution by donor T lymphocytes in the ADA-deficient mice indicates that restoration of immune function occurred by rescue of endogenous ADA-deficient lymphocytes through cross-correction from the engrafted ADA-replete donor cells. Thus, ADA-deficient SCID is unique in its responses to nonmyeloablative bone marrow transplantation, which has implications for clinical bone marrow transplantation or gene therapy

A quick and simple method, usable in the field, for collecting parasites in suitable condition for both morphological and molecular studies

11 pages impriméesInternational audienceMany methods have been proposed for collecting and fixing parasites, but most were written before the molecular age, and were intended to be practised by experienced parasitologists in well-equipped laboratories. We describe here a very simple method, illustrated by photographs, for collecting helminths from the digestive tract of vertebrates. It only requires a few plastic vials, some ethanol and a means to heat water. Basically, the method consists of: (a) the extraction of all organs from the abdominal cavity; (b) opening the digestive system longitudinally; (c) agitate gut and contents in a saline solution (i.e. ca. 9% NaCl or ¼ sea water in tap water); (d) decant in saline as many times as needed to clean contents; (e) immediately fix parasites in near-boiling saline; (f) discard saline and keep specimens in 95% ethanol. Additional information is given for collecting parasites from fish gills with a similar process. The method will collect most helminths (digeneans, larval cestodes, nematodes, acanthocephalans) from the digestive tract, and monogeneans and isopod and copepod crustaceans from fish gills. The specimens will be suitable for both morphological study and DNA sequencing. The method is simple, fast, inexpensive, and can be used by untrained personnel, even in the field without electricity and without a binocular microscope. It can also be used by trained parasitologists who need to expedite treatment of abundant samples