Demonstration of le cells

I have listed out the LE cells structure and Microscopical examinaton of LE CELLS, Difference between tart cells and le cells, clinical symptoms and diagnostic procedure.

1. Demonstration of LE cells PRESENTED BY, S.SHRUTHI VASAN III BSC.CLT Dr.NGP ARTS & SCIENCE COLLEGE

2. Le cells: An  LE cell  is a Neutrophil or Macrophage that has phagocytised (engulfed) the denatured nuclear material of another cell. The denatured material is an absorbed haematoxylin body (basophilic particle also called an LE body). They are a characteristic of lupus erythematosus , but also found in similar connective tissue disorders. The LE cell was discovered in bone marrow in 1948 by Malcolm McCallum Hargraves (1903–1982), a Physician and Practicing Histologist at the Mayo Clinic. Classically, the LE cell is analyzed microscopically, but it is also possible to investigate this phenomenon by flow cytometry .

3. FLOW CYTOMETER

4. SLE Persons having lupus erythematosus , one of the "collagen" diseases, have an abnormal plasma protein that causes swelling and breakdown of certain blood cell nuclei in vitro. This degenerated nuclear material attracts phagocytic cells, particularly segmented neutrophils , which engulf this nuclear mass. The resulting phagocyte and inclusion material is termed an "L.E." cell. Lupus erythematosus is a chronic, sometimes fatal, disease of unknown etiology . The peculiar skin eruption across the nose and cheeks (butterfly rash) and arthritis can be accompanied by various visceral manifestations. Often the rash is not present, and diagnosis depends on demonstration of the L.E. cell. Frequently the earliest symptoms appear after intense exposure to sunlight. Leukopenia, thrombocytopenia, and an elevated sedimentation rate are some of the clinical signs of the disease.

5. Two methods of demonstrating the L.E. cell and antinuclear antibodies are the Rotary bead method and Fluorescent antibody method. The rotary bead method is positive in 75-80 erythematosus . The fluorescent antibody method is positive in 95-100 patients with lupus erythematosus . The fluorescent antibody method requires equipment that limits its use to larger laboratories.

6. ROTARY BEAD METHOD Principle Leukocytes are broken down in vitro allowing the abnormal plasma protein to react on the altered nuclear material. Incubation enhances the nuclear deterioration and phagocytises. Slides are prepared and examined for the peculiar "L.E." cell.

7. Free masses of lysed nuclear material, with or without polymorphonuclear leukocytes clustered about them (rosette formation), are suggestive of the L.E. phenomenon. Observing "rosettes" should encourage the technician to repeat examinations and further search for the true "L.E." cells. A positive report should not be made without the identification of this cell. The inclusion body with the leukocyte is homogeneous and has no chromatin pattern. This feature distinguishes the true "L.E." cell from the "tart" cell (nucleophagocytosis). This latter cell contains an engulfed, damaged nucleus, usually that of a lymphocyte, which still contains a recognizable chromatin pattern and a distinct nuclear membrane.

8. Collection of sample:: Lupus erythematosus (LE) cell testing is performed using any of the following: Heparinized bone marrow Heparinized venous blood Oxalated venous blood Defibrinated venous blood Clotted venous blood LE factor and donor cells LE Factor: An Antibody found in the serum found in SLE Patients.

9. Obtaining bone marrow is usually distressing for the patient; therefore, the buffy coat from venous blood is an adequate substitute. If the equipment for buffy coat is unavailable, an untreated venous blood sample is left to clot (from 20-120 minutes) and the plasma removed. The residual clot is passed through a wire mesh and centrifuged for 5 minutes to obtain a buffy coat. This buffy coat is then smeared on glass slides to search for LE cells.  [2] The test may be performed by mixing the patient's plasma, serum, or serous effusions as a source of LE factor with bone marrow from a donor subject.

10. Considerations: The ideal temperature to perform this test is 22°C, and the process may be hastened by incubation at 37°C.

11. interpretation A lupus erythematosus (LE) cell test is considered positive when approximately 2%-30% of the cells seen on the slide in the neutrophil count are LE cells. A smear is considered positive when 10 or more characteristic LE cells are seen during a 15-minute search, associated with the presence of extracellular, amorphous, nuclear masses. The presence of LE cells indicates lupus. Negative findings on LE cell testing exclude (deny) a diagnosis of  systemic lupus erythematosus (SLE).

12. Positive reactions are also seen in  Rheumatoid arthritis Chronic hepatitis( lupoid ) Scleroderma Dermatomyositis   Polyarteritis nodosa Acquired  hemolytic anemia Hodgkin disease It may also be positive in persons taking phenylbutazone (Fever & RE) and hydralazine (BP)

13. application Lupus erythematosus (LE) cell testing was once performed to diagnose  systemic lupus erythematous  but has been replaced for this purpose by  antinuclear antibody testing .

14. THANK YOU!