Bone marrow transplantation in precocious osteopetrosis.
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Animals↗
Biomedical subjects
N W Nisbet
Publications and source records attributed to N W Nisbet.
Reduced life span of the osteoclast in osteopetrotic (mi and midi) mice.
Osteoclasts were enumerated on the parietal bones of mice carrying combinations of alleles (mi, midi and +) at the microphthalmic locus. Homozygous mutants at this locus show varying degrees of osteopetrosis due to defective bone resorption. As the severity of the bone resorption defect increased across the genotypes, there was an increase in the total number of osteoclasts and also an increase in the proportion of uninucleate osteoclasts. In parietal bones incubated for 24 h with parathyroid hormone (PTH) there was no significant difference between osteopetrotic and normal bones in terms of the number of osteoclast nuclei that had taken part in fusion. When bones were incubated for 24 h in the absence of PTH the percentage of osteoclasts remaining was correlated inversely with the severity of the bone resorption defect. Thus, we suggest that the increased proportion of uninucleate osteoclasts in these mutants results from a shortened life span, reducing the time-dependent accumulation of nuclei. This implies a reduced efficiency of uninucleate osteoclasts over multinucleate ones and we speculate on the reason for this.
Alleles↗
Evidence for osteoclast production in mixed bone cell culture.
Osteoclastlike cells (OLCs) were recognized by their morphological, histochemical, and behavioral characteristics. These characteristics were initially observed in freshly isolated osteoclasts when neonatal rat bone cells were introduced into culture. Epithelioid cells proliferated during the first week to form a confluent layer. The cells' alkaline phosphatase content and ability to produce elevated levels of cyclic adenosine monophosphate in response to parathyroid hormone (PTH) led us to tentatively describe them as osteoblastlike cells. When a second addition of neonatal bone cells was made onto this confluent layer, OLCs were produced, their numbers increasing to a maximum 7 days later and dying out 3 weeks later. Mononuclear cells with similar characteristics to these multinuclear OLCs were observed and their production and disappearance closely paralleled that of the OLCs. Fusion of OLCs was observed. The conditions under which OLCs were produced are described.
Animals↗
A study of osteoclasts on calvaria of normal and osteopetrotic (mi/mi) mice by vital staining with acridine orange.
A novel staining procedure for enumerating osteoclasts on neonatal mouse calvaria with the vital fluorescent dye acridine orange is described. It has the advantage over Barnicot's neutral-red method in that the nuclei and cytoplasm of the osteoclast are stained differentially. The osteopetrotic calvarium (mi/mi) has fewer multinucleate osteoclasts than its normal counterpart (mi/+) and they are differently distributed. The osteopetrotic calvarium has more mononucleate cells which stain like osteoclasts with acridine orange than the normal calvarium and these cells also are differently distributed. These mononuclear cells may be mononuclear osteoclasts or their precursors. These observations suggest that the defect resulting in this osteopetrosis lies with osteoclast differentiation.
Acridine Orange↗
A new allele of microphthalmia induced in the mouse: microphthalmia--defective iris (midi).
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Alleles↗
A secondary immune deficiency in the Fatty/Orl-op rat.
The response of T and B lymphocytes to the mitogens PHA and LPS was studied in the Fatty/Orl-op rat. Whereas T and B cells from op/op spleen showed no lack of responsiveness compared with that of normal (op/+) littermate rats, the response of thymic cells to the T cell mitogen was lower in op/op rats over 12 days of age compared with normal rats of the same age. Osteopetrotic rats of 10 days and younger did not show this T cell deficiency. The responsiveness of T cells from op/op lymph nodes to PHA was less than that of normal rats at 12 days and older. Thus, though splenic T and B cell populations were well maintained in the adult op/op rat despite the severe depletion of marrow cells in this mutant, the thymus and lymph node T cell populations were qualitatively or quantitatively deficient. Since osteopetrotic obliteration of the marrow cavities precedes the appearance of the immune deficiency we suggest that the immune deficiency may be caused by a failure of T cell supply to the thymus and lymph nodes.
Animals↗
Donor origin of the in vitro hematopoietic microenvironment after marrow transplantation in mice.
Bone marrow stroma from radiochimeric mice was established in culture. The polymorphic enzyme glucose phosphate isomerase (GPI) was used to determine the proportions of donor and recipient present in the original bone marrow and in cultured stroma. Bone marrow initially containing 95% donor GPI, when cultured and subsequently passaged for up to 8 weeks remained about 70% donor GPI. We conclude that many cultured stromal cells are donor derived in our radiochimeras and these are probably of hematopoietic origin.
Animals↗
John Charnley: 29 August 1911-5 August 1982.
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History, Modern 1601-↗
Failure of thymic grafts to stimulate resorption of bone in the Fatty/Orl-op rat.
An infusion of compatible normal bone marrow stimulated bone resorption in both unthymectomized and thymectomized Fatty/Orl-op osteopetrotic rats. Bone resorption was not stimulated by compatible normal thymic grafts in either unthymectomized or thymectomized mutants. It is concluded that the thymus is not fundamental in the cure of osteopetrosis in this strain of osteopetrotic rat and that the defect lies in myeloid rather than thymic regulation of osteoclastic function.
Animals↗
Osteopetrosis in the Grüneberg (mi) mouse can be cured by cultured allogeneic bone marrow.
H2 compatible allogeneic bone marrow cultured for four weeks was effective in inducing bone resorption when injected intravenously into osteopetrotic mi mouse. The active cell line or its progenitors can therefore be established in culture and is probably derived from a haematopoietic stem cell (HSC).
Animals↗
Versatile stem cells in bone marrow.
The question of whether there is a single pluripotent haemopoietic stem cell or a variety of stem cells each with a capacity for self-replication is still unresolved. Evidence from radiation-chimaeras and from patients with marrow grafts indicates that haemopoietic stem cells are not a homogeneous population. It is suggested that in marrow there is a variety of stem cells, some controlled by recognised factors and others by as yet unknown factors. It is postulated that some of these cells are pluripotent, whereas others are differentiated for a single line, so both polyphyleticists and monophyleticists may have been partly right.
Animals↗
Osteogenesis in osteopetrotic mice.
Ectopic bone arising in grafts of compatible normal intact bone marrow in microphthalmic osteopetrotic recipients was examined in the light microscope and was found to be unaffected by the deficiency that curtails resorption of primitive woven bone in osteopetrotic animals.
Animals↗
The origin of osteoclasts.
We are satisfied from studies with mi mi osteopetrotic mutant mice that osteoclasts arise from the myeloid tissue of bone marrow and not as formerly proposed from osteoprogenitor cells. Grafts of compatible normal myeloid cells cure the osteopetrosis by the substitution of the qualitatively defective osteoclasts with normal ones. Nevertheless it is still not fully clear through what cellular cascade this is effected. Current opinion would favour the pathway from pluripotent haematopoietic stem cells to circulating monocytes to tissue macrophages with ultimate fusion to form multinucleate osteoclasts. However, it is recorded that osteoclasts differ from macrophage polykaryons of inflammatory tissue not only in certain subcellular characteristics but in absence of Fc and C3 receptors. We can explain this as due to development through a specialised line of osteoclast precursors independent of conventional macrophages, if current unpublished experimental studies confirm the transfer to osteoclasts of the additional "beige" marker incorporated into grafted material.
Animals↗
Tissue repopulation during cure of osteopetrotic (mi/mi) mice using normal and defective (We/Wv) bone marrow.
Resorption of petrotic bone in osteopetrotic (mi/mi) mice was brought about by transplantation of bone marrow to X-irradiated recipients. In an attempt to learn more about the donor cell line involved in this process, both normal and defective marrow were used. The consequent repopulation of the lympho-myeloid complex was monitored by isoenzymes of glucose phosphate isomerase. The progress of normal marrow grafts was contrasted with that of a defective marrow (We/Wv). Despite the observation with We/Wv marrow showed reduced ability to form colonies in the spleen of an irradiated recipient, this marrow was as effective as normal marrow in inducing resorption of petrotic bone. The primordial stem cell for the osteoclast (haematopoietic stem cell?) is thus not a CFUS. Chimaeras with resolution of osteopetrosis by We/Wv bone marrow may exhibit erythropoiesis from residual stem cells of the host but leucocytes and platelets from the donor.
Anemia, Macrocytic↗
Bone absorption and the immune system.
The absorption of bone in Grüneberg microphthalmic mice and the op op osteopetrotic rat can be stimulated by an injection of compatible bone marrow, and complete resorption can occur. It is probable that the bone-absorbing osteoclast responsible is derived from a haematopoietic stem cell resident in the bone marrow (BM). Maintenance of the resorption depends on the survival of donor cells which has an important implication for clinical therapy in man. No evidence was found that the thymus played a leading role in the mechanism of bone absorption in the Grüneberg microphthalmic mouse, or in the op op rat.
Animals↗
The fate of allogeneic grafts of intact bone marrow in immunologically tolerant recipients and after abrogation of the tolerance.
In ossicles derived from grafts of compatible intact bone marrow in the subcapsular renal site the ectopic bone remains of donor provenance but the haemopoietic elements are partially replaced with tissue by host cells derived from stem cells migrating from the blood. Ossicles derived from incompatible intact bone marrow grafted in immunologically tolerant recipients are morphologically identical to those derived from compatible intact bone marrow. On abolition of the tolerance the allogeneic ectopic bone and donor microenvironment for haemopoiesis became manifest: haemopoietic marrow, even the host elements, was rapidly lost, bone disappearing rapidly if recently established, slowly if long established. Provided the state of immunological tolerance persists, the histoincompatible bone and microenvironment of the donor co-exist with marrow derived from the circulating haemopoietic stem cells of the tolerant recipient.
Animals↗
Resorption of bone.
The cell-system responsible for resorption of bone is now considered to be a derivative of haematopoietic bone-marrow, not skeletal connective tissue. Consideration of mutant mice and rats, with defects of bone resorption giving osteopetrosis, suggests that the primary defect is of the professional scavengers, the mononuclear-phagocyte system, failing to recognise effete bone. To explain associated defects of thymic lymphocytes it is postulated that the mononuclear-phagocyte system may be activated to a major or minor extent by professional recognisers, thymic lymphocytes, as happens in some inflammatory reactions.
Animals↗
Resolution and relapse of osteopetrosis in mice transplanted with myeloid tissue of variable histocompatibility.
Osteopetrotic microphthalmic mice (mi/mi) were treated by injections of suspensions of myeloid tissue, newborns i.p., and weanlings i.v. Donated syngeneic material effected permanent cure of oteopetrosis provided that the dose was large enough (10(8) cells of bone marrow). H-2-compatible allogeneic bone marrow was initially as effective, but relapse ensued in immunocompetent mice. H-2-incompatible marrow was ineffecitve except in one set of newborn tolerant mice. Total body X-radiation in sublethal doses to recipients allowed permanent cure with H-2-compatible, and, in one circumstance, with H-2-incompatible marrow in smaller doses. The best results were obtained after lethal irradiation and the smaller dose of marrow. Results were checked by chromosome assay demonstrating that cure or relapse was correlated with permanent take or rejection, respectively, of a transplant in a recipient's bone marrow. Retention of donor lymphocytes alone was not associated with effective bony resorption; the candidate cell line for effectiveness remains the haematopoietic stem cell-monocyte-tissue phagocyte.
Animals↗