Comparative Clinical Pathology

, Volume 13, Issue 2, pp 70–81 | Cite as

Haemotoxicity of chlorambucil in the Wistar Hanover rat with particular reference to bone marrow culture, marrow cell apoptosis and levels of FLT3 ligand

  • G. Molyneux
  • S. Rizzo
  • F. M. Gibson
  • S. Sulsh
  • C. M. Andrews
  • A. M. Pilling
  • T. Nakshbandi
  • E. C. Gordon-Smith
  • J. A. Turton
Original Article
First Online:
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Accepted:

Abstract

We have recently developed a new model of drug-induced chronic bone marrow aplasia (CBMA) in the mouse, which shows features of aplastic anaemia (AA) in humans. Using a regimen of repeated doses of busulphan (BU) we induced late-stage (i.e. chronic) bone marrow depression. There are reports that indicate that other antineoplastic agents [e.g. chlorambucil (CHB), mitomycin, melphalan] may also cause CBMA in the mouse. Wishing to develop a model of CBMA in the rat, we investigated the potential of CHB to induce this change. Female Wistar Hanover (Wistar Han) rats were dosed with CHB (6.3 mg/kg intraperitoneally) on six occasions over 18 days. Animals (n=6–8) were killed and sampled on nine occasions (at 1, 3, 9, 16, 24, 38, 45, 59 and 65 days) after the final CHB dose. A full blood count was performed, and serum was prepared for FLT3 analysis; marrow smears were produced, and the spleen and sternum were placed in histological fixative; femoral marrow suspensions were prepared for assessment of the nucleated cell count [femoral nucleated cell count (FNCC)], levels of apoptosis, and the clonogenic potential of the marrow [colony forming unit cells (CFU-Cs)]. Our results showed that at days 1 and 3 post-dosing, in general, red blood cells (RBCs), lymphocytes and FNCC were significantly reduced in CHB-treated animals; reticulocytes were increased, and platelets and neutrophils were unaffected. At 9 days, parameters in CHB rats were returning to normal, but lymphocytes were still decreased. At 16 and 24 days, many blood parameters were normal, except for reduced lymphocyte counts; this pattern generally remained until the end of the study (day 65). Levels of apoptosis in marrow cells of CHB-treated rats were increased immediately post-dosing, and this elevation persisted until day 16; thereafter, levels were generally normal. Serum FLT3 ligand (FL) levels showed some evidence of increases in CHB rats after dosing. CFU-Cs/femur were significantly reduced in CHB animals after dosing, returning to normal values at day 24. In general, marrow smears showed reductions in the myeloid, erythroid and lymphoid lineages of CHB animals at days 1, 3 and 9, returning to normal at 16 and 24 days. Histology of the spleen showed severe depletion of the white pulp immediately post-dosing in CHB-treated animals. Therefore, it is concluded from these findings that CHB does not induce late-stage (i.e. chronic) bone marrow aplasia in the female Wistar Han rat.

Keywords

Apoptosis CFU-C Chlorambucil FL Haemotoxicity Rat 
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Notes

Acknowledgements

We wish to acknowledge with thanks the assistance of the technical staff at the School of Pharmacy for their husbandry of the animals. We also gratefully acknowledge Janssen-Cilag Ltd., Novartis Pharma Ltd. and Amgen UK Ltd. for their kind donation of human cytokines. The Aplastic Anaemia Trust (GM, SR), The Leukaemia Research Fund (FMG), The School of Pharmacy (GM), and BIBRA (SS) supported this work.

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Copyright information

© Springer-Verlag London Limited 2004

Authors and Affiliations

  • G. Molyneux
    • 1
    • 5
  • S. Rizzo
    • 1
    • 6
  • F. M. Gibson
    • 1
  • S. Sulsh
    • 2
    • 7
  • C. M. Andrews
    • 3
  • A. M. Pilling
    • 4
    • 8
  • T. Nakshbandi
    • 5
  • E. C. Gordon-Smith
    • 1
  • J. A. Turton
    • 5
  1. 1.Department of HaematologySt George’s Hospital Medical SchoolLondonUK
  2. 2.BIBRA International Ltd.CarshaltonUK
  3. 3.Axiom Veterinary Laboratories Ltd.TeignmouthUK
  4. 4.GSK Research and DevelopmentWareUK
  5. 5.Centre for Toxicology, Department of Pharmacology, The School of PharmacyUniversity of LondonLondonUK
  6. 6.Prostate Stem Cell LaboratoryInstitute of Cancer ResearchSuttonUK
  7. 7.Microscience Ltd.WokinghamUK
  8. 8.Pathology DepartmentHuntingdon Life SciencesHuntingdonUK

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