Mar22 M3-Translational Research in Immunology

Question 1

leukemia and blood testing before 1970s

Answer 1

visual inspection by hematopathologist under the micrscope

Question 2

gold standard for dx in hematopathology today

Answer 2

cell morphology

Question 3

blood cells knowledge before 1970s

Answer 3

• lineages IDed based on morphology

- some pathological lineages identified (completely unrelated to normal)

Question 4

major change in leukemia and lymphoma characterizition as of 1970s

Answer 4

• immunology arrived so used immunophenotyping

- look at cell surface molecules and Rs by recognizing them with Ab, instead of looking at cell shape

Question 5

how leukemia charact differs in 1996 compared to 1938

Answer 5

• no importance to cell morphology
• pathological types of leukemia mixed with normal cells
• focus on surface molecules (CDs)

Question 6

method used for immunophenotyping (still used today)

Answer 6

flow cytometer and CD reagents

Question 7

how flow cytometry is interpreted (how results are red)

Answer 7

• graphs with distribution of one CD marker as a fct of another CD marker
• trained to recognize graph patterns

Question 8

problem of flow cytometry

Answer 8

can have artefacts (bad images, bad graphs), so not a perfect technique

Question 9

diff levels of evidence to align in leukemia dx

Answer 9

-clinical suspicion and then
use bx for:
-morphological dx (malignant or benign)
-immunopheno (malignant or benign)
-molecular genetics + Southern Blot (polyclonal or monoclonal)

Question 10

problem of the diff levels of evidence to align in leukemia dx

Answer 10

clinical evidence not always correlated with lab evidence (can’t say 100% it’s CDwtv negative so it’s not this cancer)

Question 11

important thing that allowed CD discoveries

Answer 11

discovery of monoclonal Abs

Question 12

state of the science before monoclonal Abs + their advantage

Answer 12

• had mouse or rabbit Abs mixtures

- monoclonal Abs good bc can make in large qts

Question 13

problem with monoclonal Abs and immunophenotyping

Answer 13

don’t know how the diff Abs compare to each other

Question 14

HLDA workshops stands for what + started where

Answer 14

• human leukocyte differentiation antigens (CDs)

- France

Question 15

goal of HLDA workshops

Answer 15

unify and standardize CD Abs and reagents nomenclature

Question 16

how HLDA workshop would work

Answer 16

• all labs send Abs to central lab that makes them react to stuff
• two Abs react to same Ag = these two Abs are the same
• give a name to that cluster of Abs that react to same Ags (CD = cluster differentiation)

Question 17

how many CDs identified now

Answer 17

over 370

Question 18

which countries are finding the most Abs

Answer 18

Canada, USA, Europe**

Question 19

organization of first HLDA workshop (Paris 1982)

Answer 19

• 3 cell lineages (B, T and M for myeloid)
• have institutions submitting Abs
• have research front labs in the middle

Question 20

why would sometimes have a lot of labs finding Ab to one CD and only one lab finding Ab to another CD

Answer 20

some CD Abs are easier to make, to find

Question 21

one of the first applications of HLDA workshop (in 2nd workshop. Boston 1984)

Answer 21

leukemia section (eliminated later. don’t want bio and clinical mixing)

Question 22

changes in 2nd HLDA workshop

Answer 22

• commercial companies added (not only labs now)
• 3 lineages (B, T and M)
• leukemia and lymphoma marker studies
• more IHC
• workshop identified CD16 to CD26

Question 23

changes in 3rd HDLA workshop

Answer 23

• CD27 to CD45 Ided
• recognition of cross-lineage markers (like CD45)
• CD in the center = comes from many labs
• middle CD = strong individual CDs.
• periphery CDs = weak individual CDs

Question 24

criteria to establish the finding of a new CD during HLDA workshop

Answer 24

need to have 2 Abs for that CD

Question 25

HLDA workshop 4 (Vienna 1989_

Answer 25

- CD46 to CD78 - core labs (leaders), intermediary labs and labs producing Abs (periphery) - use more transfectants now to ID Abs + cDNA methods - include carbohydrate epitopes

Question 26

last HLDA workshop using blind clustering (sending Ab blindly and people cluster them)

Answer 26

HLDA workshop 5 Boston 1993 (found CD79 to CD109)

Question 27

method replacing blind clustering after HLDA workshop 5

Answer 27

molecular biology

Question 28

HLDA workshop 5: periphery labs vs central labs

Answer 28

``` central = take a strong presence strategy periphery = take a specialization strategy ```

Question 29

main changes in HLDA Workshop 6 (Kobe 1996, CD106 to CD166)

Answer 29

- molecular methods mainly - debate about IC molecule (no CD name) - NO LONGER B and T cell lineages: now increasing fragmentation with cell specific Abs

Question 30

major change in HLDA workshop 7

Answer 30

- now recognize CD as the molecule (surface antigen) - no longer use CD# as the designation for the Ab - last blind workshop bc of that

Question 31

major change in HLDA workshop 8

Answer 31

- change to functional approach to find CD functions (purpose of workshops changed) - find molecule cloned and then find the Ab (instead of opposite like before)

Question 32

HLDA workshop 8 (2005) compared to earlier ones

Answer 32

- early = Abs against unknown molecules + industry providing reagents for research, dx and therapy - recent workshops = Abs made after molecule had been cloned. rather than make Ab before molecule is cloned

Question 33

change of name of HLDA workshop 8 to HCDM meaning + reflects what

Answer 33

- human cell differentiation molecule | - now any (cell surface) molecule characteristic to a differentiated cell can be the target. leukocytes don't act alone

Question 34

limitation or condition that is necessary for biomedicine

Answer 34

* need to make sure Abs and reagents used in tests are reliabe - interlab regulation - disease classif and nomenclature - practice guidelines