IMMUNOLOGY

Bios 328
a textbook-based study of immunology
Spring 2003

http://www.lehigh.edu/~sk08/Courses/Bios328/mainpage.htm
PART I

• Immnoglobulins are proteins

• Proteins are specified by genes
• There are too few genes to specify all the
antibodies.
– i. e., ~32,000 genes < 10,000,000,000 Ab’s

• How is Ig diversity specified genetically?

Ig proteins are specified by genetic
“cassettes”

• Light chains are specified by “variable” (V),

“joining (J), and “constant” (C) gene
segments (aka “cassettes”).
DNA rearrangement
and

alternative RNA spicing

Ig proteins are specified by genetic
“cassettes”

• Heavy chains are specified by “variable” (V),

“diversity” (D), “joining (J), and “constant” (C)
gene segments (aka “cassettes”).
DNA rearrangement
and
alternative RNA splicing
Another view….
PART II
• Cassettes rearrange…

• How does this happen?

• How do you get one “V” fusing to one “J” (in a light
chain)?
• In a heavy chain, a “D” fuses with a “J”; then the fused DJ
cassette fuses with a “V” cassette…

• The orderliness of this process implies that

there are genetic instructions. What are they?
Cassettes rearrange…

• The heptamer is a palindrome

– (i.e., it exhibits two-fold rotational symmetry.)

• The nonamer is AT-rich

– “Turns” refer to the DNA helix…

Cassettes rearrange…
One turn – two turn rule…

• one turn and two turn are “recombination

signal sequences”

• one turn only reacts with a two turn

• Recombination signal sequences are the

substrates of enzymes RAG-1 and RAG-2

(“RAG” = recombination-activating gene)

So… cassettes are marked by RSS
(i.e., they are substrates for recombination.)

Thus, cassettes can be fused.

What is the consequence?

Look at mouse:
A mouse has:
134 VH, 13 DH, 4 JH segments
85 V, 4 Jsegments
and 2 V, 3 J segments

Thus, a mouse has:

134  13  4 = 6968 heavy chains

85 4 = 340 kappa chain
and 2 3 = 6 lambda chains

6968 (340 + 6) = 2,410,928 antibodies

PART III (the HARD part…)

2.4 106 < 1010

So, there must be additional mechanisms of diversity

other than “fusing” “cassettes”

How does a RAG enzyme work?

Junctional flexibility
The “hairpin loop”
Junctional flexibility, “P” nucleotides,
and “N” nucleotides are added to
CDR3
Somatic hypermutation
One turn – two turn rule…
• one turn only
reacts with a two
turn

• crossover
between direct
repeats (same
transcriptional
orientation) leads
to deletion

• crossover
between indirect
repeats leads to
inversion
PART IV
• How do immunoglobulins assemble?

• Some immunoglobulins are in the surface

membrane of immature B-cells while other
immunoglobulins of the same idiotype are
secreted by mature B-cells. What’s the
difference?

• Similarly, identical variable regions can be

shared among different isotypes. How?

• B-cells are diploids with two sets of genetic

instructions. How does just one set get
expressed?