Established in 1923, the Thorndike Memorial Laboratory at Boston City Hospital was the first clinical research laboratory in a municipal hospital in the United States of America. Minor and Castle, who were the second and third directors of the Laboratory, were pioneer haematologists and clinical investigators of the highest calibre who created an atmosphere at the Laboratory that would foster patient-centred research and attract the best physician-scientists to work and train there. The haematology research division of the Laboratory made important original contributions to the understanding of the pathophysiology of anaemia, the mechanisms of red cell and platelet destruction and the phagocytic role of the spleen, the nature of haemoglobin (normal and sickle cell), the nature of haemophilia and its therapy and the early classification of lymphoma. It contributed to the Thorndike Memorial Laboratory's worldwide reputation as a model research laboratory and established its reputation as the birthplace of modern haematology.

Understanding which cells within a cancer are responsible for its initiation and propagation is vital if we are to achieve cure. If cancer stem cells are the only population able to sustain a tumour long term, designing therapeutic strategies to target this population will give medical science the best chance of long-term cure. Significant controversy remains over the existence of cancer stem cells, predominantly due to the lack of a sensitive human cancer stem cell assay. This thesis investigates whether two haematological malignancies, myelodysplastic syndromes (MDS) and chronic myelomonocytic leukaemia (CMML) can only be driven by rare and distinct cancer stem cells. We have demonstrated that low and intermediate-1 risk MDS is driven solely by the stem cell (Lin- CD34+ CD38- CD90+ CD45RA-) by developing a novel genetic approach, tracing all somatic mutations and karyotypic abnormalities back to this population. Prior to this study, very little was known about the clonal architecture of CMML. By performing detailed phenotypic, functional, molecular and genetic analysis of patients with CMML, we were able to demonstrate that the most likely candidate driver cell in these patients was also the stem cell rather than any of the down-stream progenitors. Currently, effective therapeutic strategies for MDS or CMML are very limited. Allogeneic stem cell transplantation is the only potential cure and not suitable for most patients. Cancer stem cells, including MDS stem cells are known to be highly quiescent and selectively resistant to therapy. Having demonstrated that both MDS and CMML were driven by stem cells, we developed a novel therapeutic targeting strategy. Using the thrombopoietin receptor agonist, Romiplostim, we were able to activate stem cells and enhance their subsequent sensitivity to chemotherapy dramatically. This approach may facilitate improved remission rates and prevent cancer stem cell driven relapse in many diseases.

HA22 is an immunotoxin composed of an anti-CD22 variable fragment linked to a 38 kDa truncated protein derived from Pseudomonas exotoxin A. The mechanisms of cytotoxicity and resistance of HA22 against Acute Lymphoblastic Leukaemia (ALL) and Burkitt’s lymphoma were studied. Using a bone marrow mesenchymal cell culture assay to support ALL cell viability, I? investigated the in vitro cytotoxicity of HA22 against ALL blasts from newly diagnosed and relapsed patients. There was interpatient variability in sensitivity to HA22. There was no significant difference in HA22 sensitivity between diagnosis and relapse samples but peripheral blood ALL blasts were more sensitive to HA22 than those from bone marrow. The mechanisms of resistance to HA22 were studied, using cell lines as a model. The number of CD22 sites/ cell and the rates of immunotoxin internalisation did not affect HA22 cytotoxicity. HA22 mutants with resistance to lysosomal degradation and enhanced targeting to the endoplasmic reticulum had improved cytotoxicity. The role of apoptosis pathways proteins in HA22-mediated cell death was studied. Their role is complex but raised levels of the anti-apoptotic pathway protein Bcl-2 were found in the most resistant NALM6 cell line. Penetration of HA22 into Burkitt’s lymphoma masses was studied using a flow cytometric based method. HA22 rapidly penetrated into the lymphoma masses, however a barrier to further uptake is present which could not be overcome by the addition of adriamycin or taxol in the murine xenograft model. The ability of Acute Myeloid Leukaemia (AML) blasts to create an immunosuppressive niche was investigated using a cell line model and primary patient samples. AML blasts suppress T cell proliferation through altered arginine metabolism, dependent on the enzymes arginase II and iNOS. Small molecule inhibitors to arginase and iNOS restored T cell proliferation in vitro. AML further enhances its immunosuppressive niche by transforming surrounding monocytes into an M2-immunosuppressive phenotype, in an arginase dependent manner. The immunomodulatory protein Serum Amyloid A (SAA) was secreted by AML blasts, and leads to AML chemotaxis, IL-1production, and release of S100A9 protein. Finally, invariant Natural Killer T cells (iNKT) were shown to be cytotoxic to some AML blasts, in the presence of Galactosylceramide, and thus able to restore T cell proliferation. The results provide a strong rationale for the clinical testing of these novel immunotherapeutic and immunomodulatory strategies in patients with haematological malignancies.

No abstract available. / Thesis (Ph.D.)-University of Natal, Durban, 1980.

The following blood group phenotypes and antigens were studied: Abantu , Ax, Ay, Bm, Bm-like, B3-like, "Bombay" Oh Le(a+b-), "Bombay" Oh Le(a-b-), para-Bombay, Mi(a+), Vw+, S-s-U-, Dantu, Gerbich-, P1H, STEM+, Rh :-34, Rhnu11 , Le(a-b-c-d-), McC(e+) and Wd( a+) and a new form of polyagglutination associated with haemoglobin M - type Hyde Park. The effect of inheriting a y, D--, Dc- or R1Lisa haplotype was also investigated. The following blood group antibodies were studied: anti-N in a person with type MN red cells, anti-hrs, anti-Rh34, anti-Jsb and anti-T. Type M red cells were confirmed to absorb anti-N and type N red cells not to absorb anti-M. A new technique was described for separating the two red cell populations in twin chimeras. Three XX/XX female dispermic chimeras with blood of two genetic types, two with patchy skin pigmentation, were identified. Reduced I and enhanced i antigen expression helped confirm a case of congenital dyserythropoietic anaemia type II. Oval red cells accompanying an r (dce) haplotype were found, and anti-Tja-like haemolysins were not detected in women about to abort. Aspects of haemolytic disease of the newborn due to ABO and Rh antibodies were discussed. Two new tests in which 2-mercaptoethanol was used to distinguish between IgG (7S) and IgM (19S) immunoglobulins were described. Blood group phenotype and gene frequency studies were made in Black, White, Indian and Coloured blood donors and the results were presented in 32 tables. Thirty monoclonal anti-A and 96 monoclonal antibodies for antigens in the ABO, MNSs, Rh, Lutheran, Kell, Lewis and Kidd systems and for other antigens were investigated for their activity and specificity. / Thesis (Ph.D.)-University of Natal, Durban, 1991.

To enable assessment of recombinant BFDV capsid protein (recBFDVcap) for vaccination to protect against PBFD, commercially available lovebirds (Agapornis sp.) were tested for evidence of past and current BFDV infection using PCR, HI and HA to identify suitable BFDV-free birds in which to test the vaccine. During this attempt, it was found that lovebirds from commercial aviaries were endemically infected with BFDV with evidence of up to 100% prevalence of BFDV DNA in blood samples from individual birds over time. Such an approach was abandoned as unlikely to yield suitable numbers of naïve birds to conduct a BFDV vaccination trial. As commercially available lovebirds were considered to be a poor source of BFDV-free birds, wild caught cockatoo nestlings and eggs (long-billed corella; Cacatua tenuirostris and galah; Eolophus roseicapillus) were used to assess the efficacy of BFDV vaccination using baculovirus recombinant BFDV capsid. Eggs were artificially incubated and 3 eggs successfully hatched and 1 was successfully hand-reared. All nestlings were screened for BFDV DNA in blood using PCR upon arrival then on days 11, 18 and 25 and tested for anti-BFDV antibody on the day of arrival. All hatched birds were determined to be free of BFDV DNA and BFDV HI antibody in the peripheral blood throughout the hand rearing period and the flock was considered to be suitable for a BFDV vaccination trial. Corellas (n=13) were injected with 1 mL of vaccine containing 10 μg recBFDVcap on day 0 and 0.4 mL vaccine containing 66.8 μg recBFDVcap on day 11. All vaccinated corellas and 5 non-vaccinated control corellas were given 0.4 mL BFDV suspension (titre = log2 12 HAU/50 μL) intramuscularly and 0.1 mL orally 16 days after booster vaccination. Blood was collected periodically during the vaccination period and blood and feathers collected before and after BFDV administration. Testing included BFDV DNA detection by PCR and qRT PCR (on blood) as well as serum antibody detection by haemagglutination inhibition (HI) and BFDV DNA and antigen was detected by qRT PCR and haemagglutination (HA) (on feathers), respectively. Four of 97 blood samples collected from vaccinated birds post BFDV challenge tested positive by PCR, whereas 17 of 35 samples taken from non-vaccinated control corellas tested positive. Vaccinated birds did not develop feather lesions, had only transient PCR detectable viraemia and had no evidence of persistent infection 270 days post-challenge using PCR, histopathology and immunohistochemistry (IHC). Non-vaccinated control corellas developed transient feather lesions and PCR, HI and HA test results consistent with PBFD. They were BFDV PCR positive for up to 41 days post-challenge and qRT PCR demonstrated reduced virus replication in vaccinated birds compared to non-vaccinated control birds. Thus, administration of recBFDVcap vaccine alone was found to incite an adaptive immune response in BFDV-free corellas that subsequently conferred protection against inoculation with BFDV. A commonly utilized method for excising blood dried on filter paper was proven to be of high risk of carryover contamination facilitated by a hole punch used for processing several samples. Therefore a practical method of avoiding carryover contamination was developed and used in the DNA testing procedures of the vaccination trial. Finally, the haematological characteristics of the above mentioned cockatoos were studied before and for 97 days after experimental infection with BFDV. It was found that the pre-challenge haematological values were similar between the vaccinated and non-vaccinated corellas. Most pre-challenge parameters were comparable to previously reported values of other cockatoos and psittacine birds. Significant differences were seen in both groups when comparing pre-challenge values with post challenge values for total and differential leukocyte concentrations, but PCV and TSP were not significantly affected by BFDV challenge.

Magister Scientiae (Biodiversity and Conservation Biology) / Homopus areolatus is an endemic terrestrial tortoise that resides in a Mediterranean type of climate, which is characterised by winter rainfall and mild winter temperatures. Within ectotherms, such as H. areolatus, physiological changes are elicited by changes in the ambient temperature. These physiological changes are evident in the blood profile of reptiles. I described the morphology of immature and mature erythrocytes, leukocytes as well as thrombocytes of H. areolatus. Additionally, I evaluated erythrocytes, leukocytes and thrombocytes to assess the effects of season and cohort on these cells. Blood samples were collected in 2000 and 2001 at Elandsberg Nature Reserve in the Western Cape from H. areolatus cohorts (female, male, juvenile) in all seasons (spring, summer, autumn, winter). Blood smears were made and stained with modified Giemsa stain. SigmaStat was used for all statistical analysis. Immature erythrocyte types within H. areolatus included basophilic rubricytes, polychromatophilic rubricytes and polychromatophilic erythrocytes. Upon my evaluation, I encountered evidence to suggest that small and large immature erythrocytes possibly developed from two distinctive lineages. Further research is required to discern which lineage gave rise to which immature erythrocyte type. Cohort had no effect upon immature erythrocytes. Erythropoiesis was most prevalent during winter and spring within H. areolatus. Aberrant features of erythrocytes appeared to be more prevalent during autumn, which signified the driest season with limited food and water. Mature erythrocytes play a huge role in oxygen transport and metabolism in individuals. Factors such as size and shape are relevant since small, mature, ellipsoidal erythrocytes transport oxygen more efficiently than large, spherical erythrocytes. In H. areolatus small, mature, ellipsoidal erythrocytes appeared to be most prevalent during spring and summer. During winter however, large, spherical erythrocytes appeared to be most prevalent. Thrombocytes and seven types of leukocytes were observed within H. areolatus, namely heterophils, lymphocytes, eosinophils, basophils, monocytes, plasma cells and azurophils. Among cohort and season heterophils were most prevalent overall, followed by lymphocytes and eosinophils respectively. Basophils, monocytes, plasma cells and azurophils were present but overall, were relatively few. H. areolatus appeared to be healthy, and leukocyte counts as well as its dimensions appeared to be in accordance with other reptilian studies. This study serves as the first baseline haematological reference forH. areolatus. The study forms the second of its kind on South African tortoises, only one other haematological study has been done namely, P. geometricus which is a sympatric species to H. areolatus.

No description available.

No description available.

No description available.