1. Lab 5 –Blood, Lymphatics and the Immune Response Gilbert Pitts, Ph.D., Joseph Schiller, Ph.D., James F. Thompson, Ph.D.

2. Objectives  Examine:  Lymph node slide  Lymphatic vessels on charts and models  Blood slides  Differentiate red blood cells, platelets, neutrophils, lymphocytes, monocytes, eosinophils, and basophils  Perform 2 differential WBC counts  Identify leukemia  Identify sickle cell anemia  Understand blood typing (ABO and Rh systems)  Calculate and interpret hematocrit/packed cell volume

3. The Lymphatic System  Basic organization  Lymph fluid in lymph vessels  Lymphatic organs (red bone marrow, thymus, spleen, lymph nodes, tonsils) with smaller collections of lymphatic tissue in other organs  Functions  Return interstitial fluid and proteins to the blood  Transport dietary fats to adipose tissue  Protect against cancer & infection  Lymph Flow from smallest to largest:  Capillaries  vessels  trunks  ducts  Lymph vessels anastomose and supply and drain lymph nodes along their course

4. Lymph Flow Follows Venous Circulation  Right head, arm, and thorax drained by the right lymphatic duct into right subclavian vein  Left head, arm, thorax, most of the abdominal cavity and both legs drained by the thoracic duct into the left subclavian vein R L

5.  Connective tissue capsule with trabeculae extending from cortex to medulla  Stroma – the supportive connective tissue network of reticular fibers and fibroblasts The Lymph Node capsule trabeculae

6. Lymph Node Parenchyma  Cortex - filled with lymphocytes and macrophages in follicles  Outer edge of follicle contains more T cells  Inner germinal center is the site of B-cell proliferation  Medulla - medullary cords of lymphocytes, macrophages, plasma cells (activated B cells) Cortex Medulla

7. Lymph Node Micrograph Cortex Medulla

8. Lymph Node Germinal Centers germinal centers

9.  Leukocytes = White Blood Cells  Granular leukocytes (granulocytes)  neutrophils  eosinophils  basophils  Agranular leukocytes (agranulocytes)  lymphocytes - T cells, B cells  monocytes  tissue macrophages The Formed Elements of the Blood:

10. Granular Leukocytes Neutrophil 60-70% Eosinophil 2-4% Basophil 0.5-1%

11. Agranular Leukocytes Lymphocyte 20-25% Monocyte 3-8%

12. Leukocyte Life Span and Number  5,000 - 10,000 WBC’s/mm 3 blood  RBC/WBC ratio 700/1  Differential WBC count (a standard clinical lab report)  Neutrophils 60-70%  Lymphocytes 20-25%  Monocytes 3-8%  Eosinophils 2-4%  Basophils 0.5-1%  Abnormal proportions are correlated with different types of disease processes

13. Differential WBC Count 20-25% 3-8% l 2-4% l 60-70% 0.5-1%

14. Leukocyte Identification Agranular Lymphocyte Granular Small Spherical nucleus Dark Hidden nuc. Large 2+ lobes Neutrophil Eosinophil Basophil Monocyte Red gran. Faint gran. no large granules in cytoplasm All have many large granules in cytoplasm & multilobed nuclei

15. Composition of Blood  Blood sample separates into 2 parts  plasma - straw colored liquid on top  ~55% of the volume  formed elements  ~45% of the volume red blood cells buffy coat: white blood cells and platelets

16. Hematocrit (Hct)  Packed Cell Volume is the % of the blood which is RBC’s  Males: 40-54% (47%)  Females: 38-46% (42%)  Hct indicates the status of RBC production, the state of hydration, or various disease states

17. Hematocrit Procedure

18. Blood Typing  Antigen – any substance which provokes specific immune responses  Antigenic determinants  Antigen parts which trigger the specific immune response  An antigen may be an entire microbe or only small structures such as subregions of large molecules  RBC antigens (agglutinogens) are membrane glycoproteins Most “antigens” are complex and express multiple types of antigenic determinants.

19. ABO Blood Types  2 glycoprotein agglutinogens, A & B  One gene from each parent, A, B or O  6 combinations - AA, AB, AO, BB, BO, OO (no agglutinogens)

20. ABO Blood Types  Agglutinins  Naturally occurring antibodies produced in response to the agglutinogens not present in your blood  React in antigen-antibody response to blood not of your type  blood type AB = universal recipients  blood type O = universal donors

21. ABO System

22. Rh System  Rh typing - Rhesus monkey  Those expressing Rh antigens are Rh +  Those without Rh agglutinogens are Rh -  Normally, blood does not contain Rh agglutinins  Immune system only makes agglutinins in response to specific exposure to Rh antigens  Rh sensitivity does not occur until second transfusion  Hemolytic disease of the newborn (erythroblastosis fetalis) many “blue babies” prior to WWII

23. Hemolytic Disease of the Newborn = Erythroblastosis Fetalis

24. Perform Fingerstick and Transfer Blood

25. Slide AppearanceBlood Type A B AB O

26. Please Clean Up Your Work Area  Place only lancet and capillary tubes in designated sharps containers  Place all other blood contaminated materials (gloves, alcohol wipes, paper towels, etc.) in the large red biohazard bucket at the front of the room  Place all other discards in regular trash receptacles  Disinfect your work area with the spray solution after you have concluded your blood work.

27. Homework o Complete and turn in the questions on pages 5-13 to 5-15 o Complete Assignment 5 on MasteringAandP.

28. End L ab 5