[Mesenchymal stem cells as potential source cartilage repair]. / A mesenchymalis ossejtek felhasználáisainak lehetoségei a porckárosodással járó mozgásszervi betegségek kezelésében.

Articular cartilage damaged by disease or trauma has a limited capacity for regeneration. The end stage of cartilage loss frequently leads to osteoarthritis resulting in a significantly decreased quality of life in millions of people. The surgical treatment of articular cartilage injury has always posed difficult problems for orthopedic surgeons and regarding long-term outcomes the currently available methods are unsatisfactory. The main lack of the applied methods is the appearance of the mechanically inadequate resident fibrocartilage instead of hyalin cartilage in the place of the cartilage defect. To find reliable methods for early repair of cartilage injuries seems of huge importance. Using techniques of tissue engineering, artificial cartilage fabricated in vitro has been applied for the repair and regeneration of damaged cartilage. Mesenchymal stem cells provide a source of cells for the repair of musculoskeletal tissue. Mesenchymal stem cells are multipotent cells that are capable of differentiating into cartilage, tendon, muscle, cartilage or hematopoiesis supporting marrow stroma. To ensure the successful durable integration and function of the engineered tissue requires suitable biomechanical and biochemical circumstances, and poses the challenge of handling in vitro culture of human cells, cell biology and molecular biology.

Self-renewal and differentiation of hematopoietic stem cells: a molecular approach (a review).

Two characteristics define a hematopoietic stem cell: the ability to differentiate into all hematopoietic lineages, and the ability to maintain hematopoiesis over a life span by a self-renewal process. The mechanisms that regulate the fate of blood-forming cells in vivo, however, are poorly understood. Despite the ability to culture hematopoietic progenitor cells (committed to particular lineages), in vitro culture of self-renewing multipotent stem cells has not yet been achieved. What is clear that both intrinsic and extrinsic signals regulate hematopoietic stem cell fate and some of these signals have now been identified. which will be highlighted in this review.

[B-cell content after bone marrow transplantation--molecular basis of the limited antibody diversity]. / A csontveló-átültetés után kialakuló B-sejt-készlet--a korlátozott ellenanyag-sokféleség molekuláris alapjai.

Despite B-cell counts and serum immunoglobulin levels usually normalized by one year, humoral immunity and the incidence of bacterial infections continue to be abnormal even after years following bone marrow transplantation. This immunodeficiency could be partially caused by B-cell repertoire restriction similar to that observed early in ontogeny. Immune reconstitution after haematopoietic stem cell transplantation really follows many established ontogenetic patterns relating to the appearance of particular membrane markers, immunoglobulin classes and subclasses, and onset of antigen receptor rearrangements. The sequence of events that occur during successful bone marrow transplantation can be regarded as a blueprint for immune reconstitution in other clinical settings as well. However, the repertoire does not resemble a fetal one, because it displays adult-size IgH CDR3s, adult-type immunoglobulin gene utilization and no evidence of bias towards any particular VH-gen family. Therefore, in the description and interpretation of these events, it is important to realize that immune reconstitution does not appear to recapitulate human fetal ontogeny. In terms of B lymphocyte diversity, the inadequacy of the recovering immune system is more likely to be explained by a combination of other factors--such as clonal dominance and the delayed occurrence of somatic hypermutation.

[Restricted antibody diversity after bone marrow transplantation--homogeneous immunoglobulins]. / A csontveló-átültetés után kialakuló korlátozott ellenanyagkészlet--homogén immunglobulinok.

After bone marrow transplantation, a prolonged dysregulation of humoral immunity, including restricted electrophoretic heterogeneity of serum immunoglobulins and the appearance of homogeneous immunoglobulin components, can be observed. The current study was undertaken to characterize further and define the posttransplantational incidence of monoclonal and oligoclonal immunoglobulins, as well as the clinical and laboratory correlations of these phenomena. For this purpose, serial serum protein (IgM, IgG, IgA and CRP) quantification, electrophoresis and immunofixation were performed on 29 patients undergoing allogeneic bone marrow transplantation for chronic myeloid leukemia. 23 out of the 29 patients developed transient oligoclonal and/or monoclonal gammopathies that appeared between 20 and 1750 posttransplantational days. No correlation, however, between the development of graft versus host disease, EBV or CMV infections, or any other symptoms and development of homogeneous immunoglobulin components was seen. Therefore, the development of oligoclonal and monoclonal gammopathies after bone marrow transplantation may be an ubiquitous finding reflecting the inadequacy, i.e. oligoclonality of the recovering B-cell system.

[Regeneration of the immune system after bone marrow transplantation]. / Az immunrendszer regenerációja csontvelö-átültetés után.

After haematopoietic stem cell transplantation, reconstitution of bone marrow consists of two distinct phenomena, numerical recovery of bone marrow cellular elements on the one hand and functional recovery of cellular interactions on the other. Immune reactivity during the first month postgrafting is extremely low. Cytotoxic and phagocytic functions usually recover by day 100, while more specialized and cooperative functions of T and B cells remain impaired up to one year or more postgrafting. Regeneration of total CD4+ T cell number in adult (and especially in elderly) transplant recipients is severely limited and occurs largely by peripheral expansion of mature CD4+ T cells. While restoration of total CD8+ T cell number is commonly seen in adults, potentially important alterations in the subset composition of CD8+ populations remain. Contracted T cell repertoires for CD4+ and CD8+ T cells are consistently found in adults after T cell regeneration. This suggests that thymic function is frequently limiting in adults and that thymic-independent pathways are insufficient for restoring host immunocompetence. Although there are similarities in immune reconstitution after alllo- and autologous haematopoietic stem cell transplantations, allogeneic transplantation involves graft versus host disease and the use of immunosuppressive therapy to control it, both of which further interfere in the early developmental stages of immune reconstitution.

[Impact of DNA chips on haematological oncology] / DNS-chipek alkalmazása az oncohaematologiában.

As the Human Genome Project hurtles towards completion, DNA microarray technology offers the potential to open wide new windows into the study of genome complexity. DNA chips can be used for many different purposes, most prominently to measure levels of gene expression (messenger RNA abundance) for tens of thousands of genes simultaneously. But how much of this data is useful and is some superfluous? Can array data be used to identify a handful of critical genes that will lead to a more detailed taxonomy of haematological malignancies and can this or similar array data be used to predict clinical outcome? It is still too early to predict what the ultimate impact of DNA chips will be on our understanding of cancer biology. There are many critically important questions about this new field that are yet unaddressed. By the publication of this article, it is hoped that the technology of DNA chips will be opened up and demystified, and that additional opportunities for creative exploration will be catalysed.

[A stem cell ... is stem cell?]. / Egy óssejt ... az egy óssejt?

We are used to associate stem cells with renewable tissues such as blood, gut and skin. But some cells in the adult central nervous system have the capacity to generate new neurons and glial cells as well and as such, they are considered to be neural stem cell. Yet their ability to generate neurons and glia, and their presence in the central nervous system throughout life, suggests new, intriguing possibilities for recovery and repair after damage to the central nervous system--and unexpectedly, the regeneration of blood tissues. After transplantation into irradiated hosts, neural stem cells were found to produce a variety of blood cell types including myeloid and lymphoid cells as well as early hematopoietic cells. Therefore, the developmental potential of stem cells is not restricted to the differentiated elements of the tissue in which they reside. Multipotential stem cells can persist in an undifferentiated state, and depending on specific environmental conditions function as a stem cell for many different tissues.

[The origin, development and aging of hematopoietic stem cells]. / A haematopoeticus óssejtek eredete, fejlódése és öregedése.

The past few years provided a number of challenges to our expectations regarding hematopoietic stem cell biology. Evidence has emerged that hematopoietic stem cells arise intraembryonally before they can be detected in the yolk sac. A number of genes that may regulate the formation, self-renewal, or differentiation of stem cells have been identified. Although different groups have attributed different properties to hematopoietic stem cells, it now appears that the differences may be explained by the existence of various stem cell populations, each with different potencies ranging from totipotent to more specialized, exist and may persist into adult life. Finally, we propose that the number of hematopoietic stem cells available to an individual is finite and the 'quality', although currently a subjective parameter, is of newly appreciated importance.

Beneficial effect of a human monoclonal IgM cryoglobulin on the autoimmune disease of New Zealand black mice.

NZB mice spontaneously develop an autoimmune disease characterized by autoimmune hemolytic anemia, thymic atrophy, lymphoid hyperplasia, and hypergammaglobulinemia. The aim of this study was to examine the hypothesis that cryoglobulins may have an immunoregulatory effect on the autoimmune process. The effect of human monoclonal IgM cryoglobulin preparations (including Cryo13, Cryo14, and Cryo16) isolated from the serum of patients with Waldenström's macroglobulinemia on the autoimmune disease of NZB mice was therefore studied. The effect of cryoglobulin preparations was evaluated on several disease parameters, i.e., survival, severity of anemia, and serum IgM and IgG levels (hypergammaglobulinemia). We found that immunization of NZB mice with Cryo13 at 3 months of age delayed the course of the disease, whereas Cryo14 and Cryo16 were ineffective. Furthermore, the effect of Cryo13 was long lasting. On the other hand, Cryo13 was able to react with 8 of 32 mouse monoclonal natural IgM autoantibodies. In contrast, Cryo14 was able to bind only 2 and Cryo16 none of these mouse monoclonal IgM antibodies. These results indicate that, in this model of autoimmune pathology, the beneficial effect of Cryo13 is mediated by its idiotypic interaction with the murine natural autoantibody network.

Partial characterization of two lymphocyte-specific natural autoantibodies isolated from newborn mice.

The early antibody-repertoire is mainly composed of multi- and polyreactive IgM autoantibodies. Some of them (about 10%) also react with cell surface antigens. It is supposed that these latter antibodies promote--particularly under the maturation of the immune system--the expansion of different lymphocyte populations and subpopulations, namely the filling of the secondary lymphoid organs by immunocompetent cells. In order to verify this hypothesis we established hybridoma clones from newborn Balb/c mice. Almost all IgM antibodies produced by these hybridomas (94%) were found to be polyreactive, but only two--the 4/5 and the 8/8--react with cell surface structures. The 4/5 IgM recognizes a glicerin-phosphathydil-inositol (GPI)-anchored membrane component expressed on the early thymocytes. The 8/8 antibody reacts with resting splenic B lymphocytes as well as B and T cell blasts via an about 40 kD protein. They do not influence the spontaneous or the mitogen induced proliferation of the lymphocytes. Therefore we cannot confirm the hypothesis that increase of the expansion of peripheral lymphocytes would be the main function of these neonatal natural autoantibodies.

Comparison of the lymphoid toxicities of mitobronitol and busulphan in mice: reduced B cell toxicity and improved thymic recovery as possible contributors to the reduced risk for complications following BMT with mitobronitol preconditioning.

It has previously been reported that the use of mitobronitol (dibromomannitol, DBM) instead of busulphan (BU) for myelosuppression is associated with significantly decreased risk for several complications of allogeneic bone marrow transplantation in accelerated chronic granulocytic leukemia. In exploring the pharmacologic basis for this observation, we have compared the acute and subacute cytotoxicities of DBM and BU on the spleen and thymus of mice. While there was comparable early (day 3) weight loss in both organs following these treatments, splenic B cells exhibited significantly less damage, and thymic regeneration (over weeks) was significantly faster following DBM treatment than with BU. These observations raise the possibility that improved post-BMT immune recovery could contribute to the clinical benefits observed with DBM-preconditioning.

[Immunologic bases of graft vs. host disease: 1997]. / A graft versus host betegség immunológiai alapjai: 1997.

The development of a graft-versus-host reaction after hematopoietic stem cell transplantation is an important determinant of the transplant outcome. The existence of subsets of CD4+ helper T-lymphocytes that differ in their cytokine secretion patterns and effector functions provides a framework for better understanding and improved management of the graft-versus-host reaction. It should allow improved prevention and treatment of graft-versus-host disease and the development of new strategies to enhance a graft-versus-leukemic relapse after transplantation.

Production of cytokines by rat immunocytoma clonal variants: a new type of intratumor heterogeneity.

A long-term tissue culture line of highly metastatic IR202 immunocytoma of LOU rats, and five of its clones (B4, C2, C4, C5, D3) was examined for their ability to produce cytokines. A combination of cytokine detection bioassays was employed in order to compensate for differences in sensitivity and specificity, and the possibility of inhibitors masking an activity. All the IR202 variants tested were shown to secrete constitutively varying amount of interleukin (IL-6). In addition, most of the cells produce IL-10 and TGF-beta, except clones C4 and C2, respectively. Moreover, supernatants from clone B4, C2, C4, and D3 may contain low levels of soluble IL-1. Membrane-bound IL-1 was found on the surface of B4, C4, and D3 cells. None of the IR202 variants, however, produced IL-2, IL-4, tumor necrosis factor-alpha (TNF-alpha), or LT. These results are consistent with the concept of a new type of intratumor heterogeneity and the ability of immunocytoma tumors to generate different immunoregulatory molecules in tumor-bearing hosts.

Functional heterogeneity of in vitro selected variants from an IgM-secreting rat immunocytoma.

A long-term tissue culture line of highly metastatic IR202 immunocytoma of LOU rats, and five of its clones (B4, C2, C4, C5, and D3) were established and studied comparatively. All such cells were similar in terms of: (i) light microscopic morphology, (ii) growth rate, (iii) saturation density, (iv) cell cycle progression, and (v) cell surface IgM, major histocompatibility complex (MHC) class I and class II antigen expression, but (vi) showed a non-homogeneous pattern of chromosomal constitution, with both numerical and structural abnormalities detected in variable proportions of the different cell variants. Moreover, IR202 variants exhibited a marked difference in the production of soluble factors which was closely associated with the ability of their supernatants to inhibit mitogen (affinity-purified goat F(ab')2 fragments specific for rat mu-chains (anti-mu antibody), lipopoly saccharide (LPS), or concanavalin A (ConA))-induced proliferation of normal splenic B and/or T lymphocytes. These results are consistent with the concept of intratumor heterogeneity and the ability of immunoglobulin-secreting tumors to induce severe immune dysfunction in host animals and humans.

A natural IgM antibody does inhibit polyclonal and antigen-specific IgM but not IgG B-cell responses.

Since a B-cell growth-inhibitory natural IgM antibody was identified in the culture supernatants of LPS-stimulated murine splenic B lymphocytes [11], attempts have been made to define other possible functional role(s) of this antibody. Here we show that this regulatory IgM is able to inhibit not only the proliferation of splenic B cells, but also their IgM secretion during LPS-induced polyclonal, as well as antigen (FITC-KLH)-specific antibody responses. In contrast, IgG1 production of hapten (FITC)-specific B cells neither during restimulation with LPS nor in the presence of carrier-specific T lymphocytes in vitro was affected by regulatory IgM. Therefore, whereas newly emerging naive B cells are highly susceptible, IgG-secreting B cells appear to be completely resistant to inactivation by the regulatory IgM autoantibody.

Novel regulators of the humoral immune response.

Activation of mature B cells to proliferation and terminal differentiation is a multistep process controlled mainly by macrophages and T cells. However, there is growing evidence that B cells and other factors can also regulate the humoral immune response. Here, Ferenc Uher, Eva Rajnavölgyi and Anna Erdei describe their work on the emerging role of regulatory interactions between subsets of B cells and their soluble products.

Down-regulation of murine B lymphocyte growth: arrest of B cells in G1 underlies immunosuppression induced by an IgM antibody.

Previous studies have demonstrated that culture supernatants of LPS-stimulated murine splenic B lymphocytes (BLPSSN) are able to inhibit the growth of freshly isolated B cells via an IgM antibody. In this work we investigated the progress of LPS-activated B lymphocytes through the cell cycle in the presence of this antibody. We found that the regulatory IgM did not affect the entry of LPS-stimulated B lymphocytes into G0*, as assessed by the increased expression of I-A antigens. Events that characterize the early G1 phase (G1A), such as cell enlargement and increased RNA synthesis, also occurred in the presence of the antibody. In contrast, events which mark the G1B phase, such as further cell enlargement and late RNA synthesis were inhibited. Moreover, a significant portion of the cells failed to incorporate [3H]thymidine and did not progress through S, G2, or M, as revealed by their DNA content. Therefore, our work points toward a well-defined stage of the early G1 phase at which the antibody inhibits the progression of B lymphocyte activation. This result shows a new insight into the mechanism of antibody-mediated down-regulation of polyclonal B cell responses.

Autocrine regulation of murine B lymphocyte growth by an IgM antibody.

Culture supernatants of LPS-stimulated murine B lymphocytes are able to inhibit the growth of freshly isolated splenic B cells via an IgM antibody. The binding specificity of this IgM is not yet defined, but appears to be a B lymphocyte surface structure distinct from membrane immunoglobulin, MHC class II antigen, transferrin and Fc gamma receptors, and B220. The regulatory autoantibody allows the normal progression of early, but not late steps in the cycle of polyclonally-stimulated B lymphocytes and does not affect the increased antigen-presenting capacity of activated B cells. Therefore, this autoregulatory cycle is apparently ubiquitous and may be a major component of B lymphocyte homeostasis under physiological, as well as pathological conditions. Moreover, these findings bring into focus a possible regulating role of B lymphocytes in the humoral immune response.