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Haemoglobin Westmead, α2122(H5)GInβ2: A new haemoglobin variant with the substitution in the α1β1 contact area
314
HSA AND ASBT
Pathology (1980), 12, April
HAEMOGLOBIN WESTMEAD, a21221H5) G'"p2:A NEW HAEMOGLOBIN VARIANT WITH THE SUBSTITUTION IN THE alp, CONTACT AREA
P. J . FLEMING, W. G. HUGHES,R. K. FARMILO, K. WYATT & W. N. COOPER Haematology Department, Institute of Clinical Pathology and Medical Research, Westmead, New South Waies A new haemoglobin variant, Haemoglobin Westmead a21?2(H5)C'np,, is described. The variant was detected in a 30yr-old Chinese female who, at first presentation, was diagnosed as having iron deficiency anaemia. The anaemia responded to iron therapy, however, abnormal red cell changes were still evident. The possibility of a haemoglobinopathy was then investigated. No electrophoretic evidence of a haemoglobinopathy could be found. The haemoglobin A, and foetal haemoglobin levels were normal. On supravital staining with brilliant cresyl blue, occasional haemoglobin H cells were demonstrated. After overnight incubation with new methylene blue stain, inclusions were present in all red cells. No increased heat instability was demonstrated. These findings suggested the possibility of a thalassaemia trait and a haemoglobin variant. By globin electrophoresis and column chromatography a variant a chain was separated and analysis of the variant c( chain indicated a substitution of a glutamine residue for a histidine residue at 122 of the ct chain. As the histidine residue at 122 is involved in the alpI contact in the normal haemoglobin molecule, a substitution could be expected to disrupt the structure of the haemoglobin molecule and lead to instability. In the case of Haemoglobin Westmead, the effect on stability appears to be minimal.
THE MECHANISM OF IMMUNE SUPPRESSION IN BALBIC MICE BEARING THE PLASMACYTOMA TEPC-183
D. E. JOSHUA*, G. BROW@& I. C. M. M A C L E N N A N *Haematology ~ Department. Royal Prince Alfred Hospital, Sydney and tNuj?eld Department of Medicine, Radclgfe InJirmary, Oxford, U . K . Mice bearing transplantable plasmacytomas have an immune defect closely resembling the defect of patients with multiple myeloma. They have a severe impairment of the primary immune response to both T-dependent and Tindependent antigens, yet their secondary immune response is much less impaired. The aim of this series of experiments was to document the primary immune response of mice bearing the plasmacytoma TEPC-183. Healthy mice and mice bearing the reported non-immunosuppressive tumour MOPC104E were used as controls. The response to chicken red blood cells (CRBC) was documented by a complement dependent cytotoxicity assay for IgM and an antibody dependent cellular cytotoxicity assay for IgG. The defect was shown to affect both the primary IgM and IgG responses to CRBC but could be overcome either by increasing the antigen dose or by using Freund's complete adjuvant together with antigen. Immunological memory was also impaired in TEPC-183-bearing mice. In a second series of experiments the primary immune responses of immunologically deprived syngeneic mice were measured after they had been reconstituted with cells from normal or tumour-bearing mice. Lymphocyte reconstitution experiments were carried out in mice which had been irradiated with 950 R. Lymphoid preparations from TEPC- 183-bearing mice were as effective as those from healthy controls in reconstituting primary immune responses. When mice were deprived of macrophage function using large doses of horse red blood cells, macrophage preparations from normal mice were able to restore primary immune responsiveness partially. However, macrophages from TEPC-183-bearing mice were unable to bring about such restoration. It is concluded that the impairment of the primary immune response of mice bearing the plasmacytoma TEPC183 is due to a macrophage rather than lymphocyte abnormality.
EFFECTS OF COLCHICINE AND VlNCRlSTlNE ON POLYMORPHONUCLEAR LEUCOCYTE FUNCTION IN VlTRO
M. M. WOLF,M. A. QUINN& I. A. COOPER Haematology Research Unit, Cancer Institute, Melbourne The effect of the drugs colchicine and vincristine on normal polymorphonuclear leucocytes (PMNs) was investigated in vitro. These drugs are known to interfere with the organization of microtubules both in the mitotic spindle of dividing cells, causing metaphase arrest, and in the cytoplasm of other non-dividing cells affecting a number of normal cellular functions. PMNs were obtained from healthy volunteers by centrifugation of
HSA AND ASBT
Pathology (1980), 12, April
HAEMOGLOBIN WESTMEAD, a21221H5) G'"p2:A NEW HAEMOGLOBIN VARIANT WITH THE SUBSTITUTION IN THE alp, CONTACT AREA
P. J . FLEMING, W. G. HUGHES,R. K. FARMILO, K. WYATT & W. N. COOPER Haematology Department, Institute of Clinical Pathology and Medical Research, Westmead, New South Waies A new haemoglobin variant, Haemoglobin Westmead a21?2(H5)C'np,, is described. The variant was detected in a 30yr-old Chinese female who, at first presentation, was diagnosed as having iron deficiency anaemia. The anaemia responded to iron therapy, however, abnormal red cell changes were still evident. The possibility of a haemoglobinopathy was then investigated. No electrophoretic evidence of a haemoglobinopathy could be found. The haemoglobin A, and foetal haemoglobin levels were normal. On supravital staining with brilliant cresyl blue, occasional haemoglobin H cells were demonstrated. After overnight incubation with new methylene blue stain, inclusions were present in all red cells. No increased heat instability was demonstrated. These findings suggested the possibility of a thalassaemia trait and a haemoglobin variant. By globin electrophoresis and column chromatography a variant a chain was separated and analysis of the variant c( chain indicated a substitution of a glutamine residue for a histidine residue at 122 of the ct chain. As the histidine residue at 122 is involved in the alpI contact in the normal haemoglobin molecule, a substitution could be expected to disrupt the structure of the haemoglobin molecule and lead to instability. In the case of Haemoglobin Westmead, the effect on stability appears to be minimal.
THE MECHANISM OF IMMUNE SUPPRESSION IN BALBIC MICE BEARING THE PLASMACYTOMA TEPC-183
D. E. JOSHUA*, G. BROW@& I. C. M. M A C L E N N A N *Haematology ~ Department. Royal Prince Alfred Hospital, Sydney and tNuj?eld Department of Medicine, Radclgfe InJirmary, Oxford, U . K . Mice bearing transplantable plasmacytomas have an immune defect closely resembling the defect of patients with multiple myeloma. They have a severe impairment of the primary immune response to both T-dependent and Tindependent antigens, yet their secondary immune response is much less impaired. The aim of this series of experiments was to document the primary immune response of mice bearing the plasmacytoma TEPC-183. Healthy mice and mice bearing the reported non-immunosuppressive tumour MOPC104E were used as controls. The response to chicken red blood cells (CRBC) was documented by a complement dependent cytotoxicity assay for IgM and an antibody dependent cellular cytotoxicity assay for IgG. The defect was shown to affect both the primary IgM and IgG responses to CRBC but could be overcome either by increasing the antigen dose or by using Freund's complete adjuvant together with antigen. Immunological memory was also impaired in TEPC-183-bearing mice. In a second series of experiments the primary immune responses of immunologically deprived syngeneic mice were measured after they had been reconstituted with cells from normal or tumour-bearing mice. Lymphocyte reconstitution experiments were carried out in mice which had been irradiated with 950 R. Lymphoid preparations from TEPC- 183-bearing mice were as effective as those from healthy controls in reconstituting primary immune responses. When mice were deprived of macrophage function using large doses of horse red blood cells, macrophage preparations from normal mice were able to restore primary immune responsiveness partially. However, macrophages from TEPC-183-bearing mice were unable to bring about such restoration. It is concluded that the impairment of the primary immune response of mice bearing the plasmacytoma TEPC183 is due to a macrophage rather than lymphocyte abnormality.
EFFECTS OF COLCHICINE AND VlNCRlSTlNE ON POLYMORPHONUCLEAR LEUCOCYTE FUNCTION IN VlTRO
M. M. WOLF,M. A. QUINN& I. A. COOPER Haematology Research Unit, Cancer Institute, Melbourne The effect of the drugs colchicine and vincristine on normal polymorphonuclear leucocytes (PMNs) was investigated in vitro. These drugs are known to interfere with the organization of microtubules both in the mitotic spindle of dividing cells, causing metaphase arrest, and in the cytoplasm of other non-dividing cells affecting a number of normal cellular functions. PMNs were obtained from healthy volunteers by centrifugation of