nr probówki

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1 Name and surname Date Class 1 Topic: Humoral innate immunity 1. Radial immunodiffusion Describe the method Interpret the outcome using the reference tables. mm = g/l (Choose the component: C3 / C4 /C1q) % hemolysis test Describe and illustrate the method: nr probówki Description: Application 3. Whole blood smear performance according to the teacher s tips. Teacher s approval

2 Name. Date 1. Peripheral blond smear - May-Grünwald and Giemsa staining Ready to use May-Grünwald stain pour over the blood smear for 5 min Rinse by water Previously prepared Giemsa stain (1 volume Giemsa + 9 volumes water) pour over the blood smear for 15 min Rinse by water Dry the blood smear and observe under microscope with 100-fold magnification using immersion oil. Do not wipe the preparations Illustrate the different types of cells identified in preparation: Neutrophil Eosinophil Basophil Lymphocyte Monocyte 2. Agarose plate method Describe the principle of the method: Application of the test: 3. Boyden s method - Describe the principle of the method Application of the test: 4. Phagocytic test by Wright Describe the principle of the method Illustrate the positive result: Fill the normal range: Percentage of phagocytic cells -.. Phagocytic index. Calculate the phagocytic index for example 5 phagocytic cells - Application of the test 5. NBT test induced and spontaneous: Describe the principle of the method: Illustrate the positive result: NBTs NBTi Fill the normal range: NBTi -.. Application of the test NBTs -. Teacher s signature

3 Class 3- Cellular adaptive immunity 1. Lymphocyte isolation of density gradient Add about 3 ml Gradisol L (about 1/3 of the tube) to the large 10 ml tube Apply the patient's whole blood (using a pipette) to the tube with Gradisol L inclined at 45 C. Do not mix the suspensions Centrifuge the tube - 15 min at 1700 rpm Download using a pipette resulting buffy coat cells and move to a clean tube Illustrate the result obtained in the tube after centrifugation: 2. Rosette assay Test principle: Type E Type EA Type EAC Estimate the preparation than illustrate and describe a positive result of the test: Application of the test- 3. Blastic transformation assay Test principle: Estimate the preparation than illustrate and describe a positive result of the test: Application of the test: 4. MIF assay Test principle: Application of the test:

4 5. Direct immunofluorescence Principles: 6. Flow cytometry Principles:

5 Class 4- Humoral adaptive immunity Lysis test Put 2 drops of sheep whole blood into a 5 ml test-tube, add 4 drops of amboceptor (rabbit antibodies) and 8 drops of complement Hold the tube in your hand for about 1-2minutes Application

6 Immunological diagnostic assay Agglutination assay 1. Agglutination platelet assay Negative control with NaCl The tested strain Result: positive/negative (underline correct) positive/negative (underline correct) Type of the assay: qualitative/semiquantitative/quantitative Test principle: (underline correct) 2. Widal assay Type of the assay: qualitative/semiquantitative/quantitative Test principle: (underline correct) Illustrate positive result: Flagellum agglutination (H) Somatic agglutination(o)

7 3. Hemagglutination assay Hemagglutination with human blood Hemagglutination with sheep blood Result: positive/negative (underline correct)) positive/negative (underline correct) Type of the assay: qualitative/semiquantitative/quantitative Test principle: (underline correct) Precipitation assay 4. Ring precipitation assay Result: Type of the assay: qualitative/semiquantitative/quantitative Test principle: positive/negative (underline correct) Other (underline correct) 5. Lysis test to small (5 ml) tube using a pipette add 2 drops of whole blood sheep, 4 drops amboceptor (rabbit antibodies) and 8 drops of complement The tube warm 1-2 minutes in your hand

8 Class 6- Immunity against infection I stage of the diagnostic procedure: 1. ELISA Sandwich type ELISA screeing test Test type: qualitative/semiquantitative/quantitative (underline the correct) Test principles: 2. ELISA - intermediate Test type: qualitative/semiquantitative/quantitative (underline the correct) Test principles: 3. ELISA - competitive Test type: qualitative/semiquantitative/quantitative (underline the correct) Test principles: IFA screening test 1.IFA -direct Test type: qualitative/semiquantitative/quantitative (underline the correct) Test principles:

9 2. IFA - indirect Test type: qualitative/semiquantitative/quantitative (underline the correct) Test principles: II stage of the diagnostic procedure: 1. Westernblot (WB) Westernblot confirmation test Test type: qualitative/semiquantitative/quantitative (underline the correct) Test principles: IFA images assessment please illustrate: Chlamydia trachomatis Chlamydia pneumoniae Mycoplasma pneumoniae Test type: direct/ indirect qualitative/semiquantitative/quantit ative Ab class: IgM/IgG/IgA Helicobacter pylori Test type: direct/ indirect qualitative/semiquantitative/quantit ative Ab class: IgM/IgG/IgA Test type: direct/ indirect qualitative/semiquantitative/quantit ative Ab class: IgM/IgG/IgA Borrelia burdgorferi Test type: direct/ indirect qualitative/semiquantitative/quantit ative Ab class: IgM/IgG/IgA Test type: direct/ indirect qualitative/semiquantitative/quantita tive Ab class: IgM/IgG/IgA IgM IgG IgA Outcomes interpretation

10 ELISA and WB assessment Helicobacter pylori Borrelia burdgorferi I stage: Test: IFA / ELISA (choose correct) IgG -.. I stage: Test: IFA / ELISA (choose correct) IgM -.. OUTCOMES SET 1 IgA.. IgG -. IgA - IgG.. IgM -. IgG - I stage: Test: IFA / ELISA (choose correct) IgG -.. I stage: Test: IFA / ELISA (choose correct) IgM -.. OUTCOMES SET 2 IgA.. IgG -. IgA - IgG.. IgM -. IgG -

11 I stage: Test: IFA / ELISA (choose correct) IgG -.. I stage: Test: IFA / ELISA (choose correct) IgM -.. OUTCOMES SET 3 IgA.. IgG -. IgA - IgG.. IgM -. IgG - I stage: Test: IFA / ELISA (choose correct) IgG -.. I stage: Test: IFA / ELISA (choose correct) IgM -.. OTCOMES SET 4 IgA.. IgG -. IgA - IgG.. IgM -. IgG -

12 Class 7- Hypersensitivities I. Early-type hypersensitivity (type I) describe a) Examples of diseases b) Describe diagnostic algorithm: Anamnese Blood morphology assessment Total IgE level assessment (test RIST) Specific IgE level assessment (test RAST) Skin tests Provocation tests Other II. Cytoctoxic hypersensitivity mechanism (typ II) describe a) Examples of diseases: b) Describe diagnostics protocols Direct Coombs reaction-

13 Indirect Coombs reaction Other: III. Immune complexes related hypersensitivity (typ III) describe a) Examples of diseases: b) Describe diagnostics protocols: IV. Delayed type hypersensitivity (typ IV) describe a) Examples of diseases b) Diagnostic protocols: Skin test Tuberculine reaction -

14 Topic: Autoimmune diseases IFA assessment please illustrate: ANA ANA ANA Test: direct/indirect Klasa przeciwciał: IgM/IgG/IgA Substrat: Typ świecenia: Test: direct/indirect Klasa przeciwciał: IgM/IgG/IgA Substrat: Typ świecenia: Test: direct/indirect Klasa przeciwciał: IgM/IgG/IgA Substrat: Typ świecenia: ASMA AMA ANCA Test: direct/indirect Ab class: IgM/IgG/IgA Substrate: Luminescence type: Test: direct/indirect Ab class: IgM/IgG/IgA Substrate: Luminescence type: Test: direct/indirect Quantitative/semiquantitative/qualitative Quantitative/semiquantitative/qualitative Quantitative/semiquantitative/qualitative Quantitative/semiquantitative/qualitative Quantitative/semiquantitative/qualitative Quantitative/semiquantitative/qualitative Ab class: IgM/IgG/IgA Substrate: Luminescence type:

15 OUTCOMES SET 1 AND 2 IFA and WB outcomes assessment please interpret: ANA ANA I stage: Test: IFA IgG ratio -.. Luminescence type: I stage: Test: IFA IgG ratio -.. Luminescence type: IgG -. Positive/border line/negative IgG -. Positive/border line/negative

16 Class 10- Transplant immunology and graft rejection 1. Please describe: General principles: Lymphocytotoxic test PRA Cross-match Lymphocytotoxic test Principles: Principles: Principles:

17 2. Please illustrate positive and negative outcome of lymphocytotoxic test Negative Positive Terasaki plate 3. Please interprete HLA assessment performed with molecular method PCR-SSP

18 Class 10- Transplant immunology and graft rejection- practical tasks Task 1 Unrelated kidney donor- HLA: HLA - A 1,3; B4,12; DR 1,8 Blood group: A Rh (+), Viral status : HIV (-), HCV (-), HBV (-), CMV(-) Please match the most suitable recipient: - Recipient 1: HLA A1,3: B4,8; DR 1,12 Blood group: O Rh (+), Viral status : HIV (-), HCV (-), HBV (-), CMV(-) Recipient 2: HLA A 1,2; B4,8: DR 7,12 Blood group: A Rh (+), Viral status : HIV (-), HCV (+), HBV (-), CMV(-) Recipient 3: HLA A3,32; B 4,8; DR1,8, Blood group: A Rh (+) Viral status : HIV (-), HCV (-), HBV (-), CMV(-) Task 2 The parents have following HLA antigens: - mother HLA A1,3; B4,8;Cw 4,-; DR 1,12 - father - HLA A2,11; B7,12;Cw 1,5; DR 1,7 What HLA combinations can be inherited by their children? Task 3 Unrelated donor has the following antigens: HLA - A 1,3; B4,12; DR 1,8 Blood group: A Rh (+) Viral status : HIV (-), HCV (-), HBV (-), CMV(-) Recipients HLA A 2,3; B7,8; DR 1,5 Blood group: A Rh (+), Viral status : HIV (-), HCV (-), HBV (-), CMV(-) PRA 75%, no vascular access Would you recommend performing the kidney transplantation in this case? Task 4 During routine examination of HLA antigens HLA 27 has been found. What does this mean for the individual? Task 5 Patient qualified for the second kidney transplantation HLA A 2,3: B-7,8 ; DR1,5 Previous donor: HLA - A2,11: B7,12; DR 1,4 Please match the most suitable recipient: - Donor 1: HLA A2,32; B 7,15; DR 1,8 - Donor 2: HLA A3,11; B 7,8; DR 4,5