PubMed Health⌕ Search

Biomedical subjects

J T Kemshead

Publications and source records attributed to J T Kemshead.

141 records · Page 8Linked to original sources

Monoclonal antibodies defining markers with apparent selectivity for particular haemopoietic cell types may also detect antigens on cells of neural crest origin.

Monoclonal antibodies described as reacting with particular subsets of haemopoietic cells were screened against a variety of neuronal cell lines to further investigate their true specificity. While some reagents, e.g., monoclonal anti-cALL (J5), were found only reactive with haemopoietic cells, other monoclonals, e.g., BA1, BA2, NA134, OKT6, and OKT9, also bound to neuronal cells. Further investigation into the cross-reactivity of these antibodies to a variety of neural crest derived cell lines indicated that, with the exception of OKT9, differential binding patterns to different lines were obtained. This suggests that haemopoietic cell subsets and neural crest derived tissues can share similar differentiation antigens. For monoclonals OKT9 and BA2 this observation was confirmed biochemically, showing that it is not just antigenic determinants but similar molecular weight cell surface antigens that are shared between subsets of the two major cell types. A similar analysis using monoclonal antibodies raised against human neuronal cells or cell lines again indicates that some antigens appear unique to neural tissue while others are shared by haemopoietic cell subsets.

Antibodies, Monoclonal↗

Monoclonal antibody to ganglioside GQ discriminates between haemopoietic cells and infiltrating neuroblastoma tumour cells in bone marrow.

An immunological approach has been sought for the identification of minimal metastatic spread of neuroblastoma to bone marrow. Here we describe the reactivity of the monoclonal antibody A2B5 to human neuroblastoma cell lines and fresh tumour tissue. This reagent, raised against chick retinal cells, reacts with all human neuroblastoma lines assayed although quantitative differences in antigenic expression exist between cultures. Analysis of tumour cells in heavily infiltrated bone marrow aspirates indicates that only 70% of the samples reacted with A2B5, suggesting that the heterogeneity seen in the expression of antigen on cell lines is paralleled in fresh tumour material. A2B5 showed no reactivity to either a panel of human leukaemic cell lines or normal human bone marrow, although reactivity to an occasional leukaemic marrow aspirate was detected. We suggest that A2B5 could form part of a panel of monoclonal reagents necessary for detecting metastatic spread of all neuroblastoma cells to bone marrow. Such a group of reagents may be useful therapeutically in a programmed of autologous bone marrow transplantation for the removal of tumour cells prior to reinfusion of haemopoietic cells to patients receiving high-dose chemotherapy.

Antibodies, Monoclonal↗

A monoclonal antibody detecting an antigen shared by neural and granulocytic cells.

A monoclonal antibody, MI/N1, is described that reacts predominantly with fresh neuroblastoma tissue, human neuroblastoma cell lines, and cells of the myeloid lineage. Investigation of the binding of this antibody to four different neuroblastoma cell lines showed CHP 100 bound approximately 4 times more antibody than CHP 126. Only 30% of the cells in the line CHP 100 bound MI/N1 as determined by indirect immunofluorescence. Thus, both quantitative and qualitative differences in the expression of antigen recognised by MI/N1 are detected on human neuroblastoma cell lines. Inasmuch as only five of eight marrow aspirates heavily infiltrated with neuroblasts bound the monoclonal, this also suggests a heterogeneity in antigenic expression on fresh tumour cells. Absorption studies indicate that the antigen recognised by MI/N1 is present on human foetal brain and adult human cerebellum. At a dilution of 1/750, equal volumes of foetal brain and adult cerebellum absorbed out 30 and 60% of the reactivity to the human neuroblastoma cell line CHP 100. No reactivity was found towards murine neuroblastoma cells or rat brain. Expression of antigen on cells in the myeloid lineage appears dependent upon their stage of maturation, increasing as cells mature to neutrophils and eosinophils. It is suggested that the quantitative and qualitative differences seen in the expression of antigen on neuroblastoma cells may relate to their being blocked at different stages of differentiation.

Adult↗

Developments in the biology of neuroblastoma: implications for diagnosis and treatment.

There has been little improvement in the prognosis for neuroblastoma in childhood over recent years. The authors review recent investigations into the biology of the tumour and discuss their implications for diagnosis and therapy. Immunological studies suggest a heterogeneity in cell-surface antigens on different human neuroblastoma lines/fresh tumours. The reagents being developed against these antigens will play a rôle in the diagnosis of metastatic spread, and may be useful in autologous bone-marrow transplantation programmes. The occasional in vivo differentiation of the tumour is reviewed, with particular reference to in vitro studies on the differentiation of human and murine neuroblastoma cell-lines. This work may have implications for therapy if ways can be found to cause in vivo differentiation of the tumour into a benign ganglioneuroma.

Animals↗

Thymus dependence of the IgG response: role of T cells is restricted to non-specific rather than antigen-specific factors.

We have been studying the requirement for T cells in the proliferation and maturation of partially differentiated memory cells into IgG antibody secreting cells. The role of T cells in such a system can be replaced by non-antigen-specific T-cell factor(s) present in the supernatant of a 24 h mixed lymphocyte reaction (TRF) but not by antigen-specific factors released upon in vitro induction of T-helper cells with antigen. A combination of antigen-specific and non-specific factors showed no evidence of synergy between the two sets of factors and we therefore conclude that the antigen-specific factors tested play no role in the later stages of B-memory cell maturation to IgG-antibody secreting cells.

Animals↗

A simplified method for the in vitro induction of IgG antibody in collagen coated dishes.

Culture dishes precoated with thin layers of acid soluble rat tail collagen simplify conditions necessary to obtain in vitro high IgG anti-DNP responses from primed and boosted mice. In contrast to Mishell-Dutton type cultures the use of collagen precludes the need to continually rock cultures and to add nutrients. Reduction of the cell density in collagen dishes to 10(6) cells/ml is without detriment to the antibody response. Cells adhering to collagen coated dishes after 24 h incubation provide an enriched population of IgG anti-DNP precursor cells; which will mature to antibody secretion at very low cell density (10(4) -10(5) cells/ml). However, as changes in the adhesive properties of cells to collagen occur during cell maturation, attempts to adapt the technique to obtain clones of antibody secreting cells were not wholly successful.

Animals↗

Non-specific factor replaces T cells in an IgG response to soluble antigens.

The antigen and T-cell requirements for the final stages of proliferation and maturation of DNP-KLH primed and boosted mouse spleen cells into IgG antibody secreting cells have been studied in vitro. The requirement for free antigen ceases after 24-48 h in vitro. The carrier-specific T-cell requirement for triggering of activated B cells by a soluble antigen (DNP-KLH) can be replaced in T cell-depleted cultures by non-antigen specific T cell-replacing factors (TRF). However, if the carrier protein is changed, TRF restores the IgG response of T cell-depleted cultures only if antigen is presented to B cells in particulate form, e.g. on the surface of macrophages, or in the presence of small amounts of antibody against the carrier protein. Thus, direct interaction between soluble protein and B cells is not sufficient to allow TRF to effectively replace specific T cells. Since TRF must be added at the start of culture, the initiation of B-cell maturation into IgG secretion by TRF occurs during B-cell proliferation, and is followed by further proliferation before IgG antibody can be detected.

Animals↗

IgG anti-hapten antibody secretion in vitro commences after extensive precursor proliferation.

We have investigated whether the intermediate cells that arise in primed mice after boosting require further cycles of division in culture before maturation into IgG antibody secreting cells. Killing of dividing cells between days 1--4 in culture, by exposure to BUdR-uv irradiation ablated the high IgG response observed on day 5 in control cultures. After T cell removal and replacement by a soluble factor (TRF) similar results were obtained. Thus B cell division over an extended period occurs prior to the appearance of IgG secreting cells. Furthermore, autoradiography of plaques from cultures briefly exposed to [3H]thymidine before harvest showed that some antibody secreting cells were synthesizing DNA at the time of assay.

Animals↗

Degradation of abnormal proteins in Escherichia coli. Differential proteolysis in vitro of E. coli alkaline phosphatase cyanogen-bromide-cleavage products.

Escherichia coli alkaline phosphatase has been purified and modified by either carboxymethylation or treatment with cyanogen bromide (CNBr). Only the CNBr-treated protein was degradable in an E. coli cell extract. Separation of the CNBr cleavage products by gel filtration in non-denaturing conditions gave rise to a number of oligomeric complexes, of which only those of molecular weight less than approximately 29000 were degradable in E. coli cell-free extracts. Carboxymethylation of the non-degradable complexes (greater than 29000 molecular weight) resulted in the formation of some complexes of less than 29000 molecular weight: such newly formed complexes were degradable by E. coli cell-free extracts. It is suggested that E. coli cell-free extracts may contain a protease/peptidase system which is active against peptide complexes below 29000 molecular weight, but inactive against peptide oligomers of greater molecular weight.

Alkaline Phosphatase↗

Removal of tumour cells from bone marrow: an evaluation of the available techniques.

Autologous bone marrow transplantation has offered a way of increasing the dose of drugs and radiotherapy which can be used to treat patients with malignant disease in an attempt to eradicate tumour. Bone marrow is taken prior to treatment and returned to the patient to 'rescue' haemopoietic function after ablative therapy is completed. Bone marrow removed for autograft may be contaminated with tumour cells at the time of harvest, and it is undesirable to return these to patients even though there are little data available concerning the number of tumour cells necessary to reseed various malignancies. This review considers the various methods available for removing tumour cells from bone marrow destined for autologous transplantation, and evaluates their advantages and disadvantages.

Animals↗

Treatment of neoplastic meningitis by targeted radiation using (131)I-radiolabelled monoclonal antibodies. Results of responses and long term follow-up in 40 patients.

Between 1984 and 1993, monoclonal antibodies (MAbs) radiolabelled with (131)I were administered into the CSF of 52 patients with neoplastic meningitis (meningosis) with progressive disease despite active conventional therapy. Selection of MAbs was based on immunoreactivity with patients' tumour and lack of binding to normal central nervous system (CNS) tissue. Following full clinical assessment and neuro-imaging which included isotope flow study of CSF pathways, (131)I-MAb was administered via a ventricular access device, lumbar catheter or both. Radioisotope activity varied from 25 mCi to 160 mCi in adults. Dose escalation was carried out and some patients received multiple doses. Distribution of (131)I-MAb and clearance kinetics were derived from serial scintigraphy and CSF/blood sampling. Evidence of localisation to tumour was frequently observed. Toxicity was minimal and easily treated, although one death occurred, possibly due to a seizure. The best results were obtained in primitive neuroectodermal tumour (n=22), where 53% of evaluable cases had responses and 11% had stable disease, adults responding better than children. Three exceptional survivals have been recorded; one patient leads a normal life at 10 years 11 months, one case is alive and normal at 3 years, 2 months. A third case survived in good condition for 8 years. The mean survival of responders was 39 months and non-responders 4 months. In the total series, 50% of patients survived for at least one year with 2 long term survivors. CSF therapy with (131)I-MAb appears to be valuable as a single agent or when used in combination with other modalities. Results of treating leukaemia and carcinoma cases suggest that re-seeding into the CSF compartment from active systemic disease may account for early relapse in the CNS. One carcinoma case with no apparent systemic disease made a remarkable response and survival for 4 years following a single treatment. Neoplastic meningitis generally carries a dismal prognosis. The results obtained in this initial trial are sufficiently encouraging to stimulate further attempts at CSF therapy with (131)I-MAbs.

Adolescent↗

The epidermal growth factor receptor in glioblastoma: genomic amplification, protein expression, and patient survival data in a therapeutic trial.

Human glioblastomas were evaluated for overexpression of the epidermal growth factor receptor (EGFR) in a therapeutic trial with the anti-EGFR antibody EMD 55900. A total of 55 cases were examined by immunohistochemistry using 4 different monoclonal antibodies on frozen or on paraffin sections: EGFR-1 (Amersham), E 62 (Merck), E 30 (Merck), and EMD 55900 (MAB 425, Merck). Definition for inclusion in clinical trials of EMD 55900 was an immunohistochemical overexpression of grade 4+ or 3+ in a scale of 4 grades of staining quality. The use of the 4 different antibodies gave essentially equal results. In 21 cases, the immunohistochemical results were supplemented by molecular genetic analysis of EGFR amplification on the corresponding locus of chromosome 7, using frozen tissue from the same blocks after screening for vital tumor areas. Since no other material was available, the differential polymerase chain reaction technique was applied. Interferon-gamma (IFN-gamma) served as a reference gene. In a preliminary experimental series with cases of known EGFR amplification, a densitometric EGFR/IFN-gamma ratio higher than 3 was determined as indicator for amplification of the EGFR gene. With this experimental approach we were able to identify an amplified EGFR gene in 13 specimens including 2 from recurrent glioblastomas in the same patients. All of these showed an increased immunoreactivity for EGFR protein. The degree of EGFR amplification (EGFR/IFN-gamma ratio as measured by DNA densitometry) showed a positive correlation with the grade of immunohistochemical protein expression, both in regard to the fraction of positive cells and to the overall staining intensity.(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Monoclonal↗

Identification of neural crest derived cells within the cellular microenvironment of the human thymus employing a library of monoclonal antibodies raised against neuronal tissues.

The organization of optimal microenvironmental conditions within the developing thymus for lymphatic stem cell migration and their further maturation requires cellular and humoral participation of the neural crest. Recently, the immunophenotypical (IP) heterogeneity of lymphatic cells has become a scientific fact. Monoclonal antibodies (MoABs) produced against the various subpopulations of the reticulo-epithelial cells (RE) demonstrated their heterogeneity. We suggest that with a library of MoABs, raised against normal neuronal tissues, neural tumors, and a medulloblastoma cell line, including UJ13/A, UJ127.11, UJ167.11, UJ223.8, UJ308, J1153, A2B5, 215.D11, 275.G7, 282.1, antineurofilament (NF - med. m.w.), and anti-Thy-1 it is possible to recognize cells of neural crest origin within the postnatal thymic cellular microenvironment. Evidence has been collected concerning such connections between the nervous system and the thymus, such as the production of neuropeptides, oxytocin, and neurophysin by the thymus. Our immunohistochemical study was carried out on quick-frozen sections of human postnatal thymuses removed during open heart surgery, employing an indirect, alkaline phosphatase conjugated streptavidin-biotin technique. The employed MoABs reacted with the subcapsular (outer cortex) thymic nurse cells (TNCs) and with medullary RE cells, in close contact with already mature, immunocompetent T lymphocytes ready to leave the thymic microenvironment and enter the peripheral blood. The thymic medulla's strong immunoreactivity with A2B5, which binds to the GQ ganglioside, is typical for peptide secreting cells often migrated from the neural crest. A2B5+, Thy-1+ IP was demonstrated on the large TNCs. Cortical RE cells showed reactivity with UJ127.11, UJ223.8, and UJ308. Dense expression of neural crest antigens was detected in the Hassall's bodies (HBs) employing MoABs UJ223.8, UJ308, 215.D11, and 275.G7. These results suggest a neural crest origin for TNCs and for 20% to 30% of the cells of thymic microenvironment. The outer (peripheral) part of the HBs contained functionally very active RE cells. These RE cells also expressed antigens characteristic of the neural crest, detectable with MoABs UJ127.11, UJ223.8, UJ308, J1153, 215.D11, 275.G7 and A2B5.

Animals↗