Humoral Response

• Ag contact Naïve cells primary humoral

response
• lag pase : clonal selection: differentiation
• Ab secreting plasma cells
• Memory B cells: G0 phase
• Log phase :Antibody peaks: Ig M: Ig G
• Secondary response: Memory cells > naïve cells

• Original Antigenic sin: vaccination to 1 strain

better response to infection with other strain
 Humoral Response

• Naive cell : Response • Memory B cell

• 4-7 days :Lag period • 1-3 days
• 7-10 days: peak • 3-5days
• Varies depending on • 100-1000 times
Ag :Magnitude greater
• Ig M : Isotype • IgG
• TD,TI :Antigens • TD
• Lower :Ab Affinity • Higher
 Organization and expression of IG genes

• 1010 Different antibodies

• Germline theory : germline cells large Ig genes
• Somatic theory : few Ig genes :somatic mutation increases no

Dryer and Bennet

2 gene 1 polypeptide : revolutionary model
Heavy gene :14 constant : c mu /delta/gamma/epsilon/alpha
Light lambda :22 lambda 1 and lambda 2
Light kappa 2 no subtypes
Experimental evidence: Tonegawa and Hozumi
Light chain: V variable J joining C constant
Heavy chain : V variable D diversity J joining C constant
 Variable Region gene rearrangement

• Ab producing cell : isotype switching :Ag specificity

• Light chain: any V and J :variable joins to C
• Heavy chain any V combines with DJ and then to C
• RNA polymerase binds to leader sequence 5’V
• Light : kappa or gamma
• Heavy: Cmu or Cdelta
• RNA splicing:
» introns b/w leader peptide and VDJ
» Region b/w VDJ and constant
» introns b/w 2 constant regions Cmu and C delta

• Translation
• B cell with IgD and IgM
 Recombination signal sequence
• To ensure proper rearrangment
RSS :-
• Conserved palindromic sequence
• Conserved Nonamer AT rich
• Intervening sequences :12bp or 23 bp

• 12bp:1 spacer 23bp:2 turn spacer

• 1 turn joins only 2 turn

• RSS on 3’of V,3’and 5’ of D and 5of J

• Kappa:V1spacer+2spacerJ
• Lambda: V2spacer+1spacer
• Heavy :V3’ 2spacer+5’ 1spacerD+3’ 1spacer+5’spacerJ
 Enzymes in RSS
• RSS: recombinase enzymes
• RAG1/RAG2:
cleaves 1 strand of DNA
Formation of hair pin
Ds break
SS Endonuclease
Cutting of Hairpin,addition of
Pnucleotides
Terminal deoxyribonucleotidyl transferase:TdT
Addition of 15nucleotides
Ligation of coding sequences
Double strand break repair enzymes
Productive arrangement/Non Productive rearrangement
Allelic exclusion
Generation of Antibody diversity
• Multiple germ line • Combinational VJ and
VDJ genes VDJ joining
random rearrangment
1 individual
51Vh + 27 Dh+ 6Jh 51*27*6= 8262
40V kappa+5J kappa 40*5=200
30V lambda+4J lambda 30*4=120

Antibody diversity
 Junctional Flexibility
• Joining of signal sequence: precisely
and coding sequence : imprecise
=
Non productive arrangement : apoptosis
Productive arrangement : Diversity
 Antibody diversity
• P addition • N addition:15optional
Recognition of RSS • TdT
Ss break • D..TC ……TA…J….
Hairpin formation D..AG……..AT…J….
Ds break 2) D…TCGA
Generaton sites for ……….ATAT…J
addition of 3)Optional addition of N
Pnucleotides
 Antibody Diversity
• Somatic Hyper mutation
– Within somatic cells mutation : new
combination
• Association of Heavy and light genes
combination of heavy and light genes