3.68. F. 5.38. 5.36. 5.6. G. 4.3. 4.44. 4.73. G. 4.96. 4.06. 3.55. Subtypes and viral load ... p = 0.78. 4.52±4.3. 0.91±1.21 p = 0.002. 2nd Generation EIA. OD SAM. P.
CENTRE HOSPITALIER UNIVERSITAIRE FACULTÉ DE MÉDECINE ROUEN
Professeur F. Simon
Docteur JC. Plantier
22 octobre 2002
THE PRIMATE LENTIVIRUSES DIVERSITY
• Non-human primates : > 18 different species reported as siv-infected • Human : 2 types of hiv as consequence of different crossspecies transmission
INTRODUCTION : Primates Lentiviruses (1996)
HIV-O
SIVcpz
CPZANT HIV-M ( A to H) SIV AGM
SIV agm sab SIV Mnd
SIV sm
HIV-2
INTRODUCTION : Primates Lentiviruses (2000) HIV-0 : A
HIV-O : B CPZCam/ HIV-N
CPZANT SIVrcm
HIV-M (A to K)
SIVsyk SIV Mnd Type 2 SIVagm
SIV drill
SIV Mnd Type 1 SIVsun
SIV AGM sab SIVhoe
SIV sm
HIV-2
INTRODUCTION : HIV-1
GROUP M GROUP N GROUP O
3 LINEAGES
: > 306 of cases; 11 subtypes ( A to K) : only six cases so far reported : > 500 cases ; cladistic organisation
EACH HIV-1 GROUP WOULD CORRESPOND TO INDEPENDANT TRANSMISSION OF SIVCPZ FROM CHIMPANZEE TO HUMAN
INTRODUCTION : HIV-2
A B C to G
‘ SUBTYPES ‘ (or groups ??)
: West Africa, Atlantic coast : Mali, Ivory Coast : Liberia, Sierra Leone : dead-end infections
EACH HIV- 2 SUBTYPE WOULD CORRESPOND TO INDEPENDANT TRANSMISSION OF SIVsm FROM SOOTY MANGABEYS TO HUMAN
HIV variability - High replication and mutation rate of HIV combined with the genomic integration and dormancy - half -life of plasma virion < 6 hours, 10 billions virions produced each day - RT pol-encoded lacks exonuclease activity : 1 mutation occurs each replication cycle - complex of genitically related but distinguishable variants =
HIV = QUASISPECIES
HIV diversity and epidemiology
Subtype : env gene homology > 80% - subtypes are reflecting a founder effect in a particular population like subtype B in IVDU , F in Romanian children, G in Russia - no clear founder effect in Africa / predominance of the heterosexual transmission ? - frequency of the circulating recombinant forms (CRF) 50% of the strains sequenced in Gabon in 2000 are CRF. Epidemiological advantage ??
A new strategy for Siv/hiv characterization Anti-hiv/siv antibodies detection using Synthetic peptides in 2 eias :
- First eia : highly sensitive Gp 41 id from hiv-m/n/o, sivcpz/agm/mnd/sm…
- Second eia : highly specific
V3 loop from hiv-m/n/o, sivcpz/agm/mnd/sm… Each sample is tested by at least 16 peptides
Reactive samples : RT-PCR and amplification using degenerated primers and pol sequencing
Hiv and siv detection and differenciation by synthetic peptides
peptides V3 or gp41 M HIV-1
O N cpz-gab
SIV
cpz-ant mnd agm sm
Specificity of V3 loop peptide-based EIAs
group M sera (n=142)
M
O
N
Cpz
Mnd Agm
Sm
group N sera (n=6)
M
O
N
Cpz
Mnd Agm
Sm
N
Cpz
Mnd Agm
Sm
HIV-2 sera (n=150)
group O sera (n=56)
M
O
M
O
N
Cpz
Mnd Agm
Sm
PCR strategy ; HIV and SIV pol gene amplification POL CAM 50 450 LENTIPOL S ET AS 1020
UNIPOL3 1940
POL CAM 52 POL LUP S 1120 POL LUP AS 900 1020
MP2
MP5
UNIPOL1 2420
UNIPOL 2 2700
H POL 4235 2570
MP3
UNIPOL4 2900 H POL 4538
H POL 4327 2650
H POL 4481 2800
MP6
More than 10 sets of primers (including highly de generated primers) needed to successfully amplified the 3 ’ pol
HIV-1 DIVERSITY SURVEY IN FRANCE A NATIONAL ANRS NETWORK
17% of non-B infection in France 25 % of non-B in Paris area All the subtypes ( A to K) are represented Importance of the heterosexual transmission
HIV-1 diversity survey in france A national anrs network
• subtype B : still predominant in homo/bisexual and IVDU • incidence of non-B in France < 1996 : 10 % >1996: 21 % • HIV-1 non B 46% of heterosexual infection 90% of African patients /partners •frequency of the circulating recombinant form (CRF) in non-B •report of gag non-B and env- B CRF
Subtypes and viral load
106
Monitor/Monitor 1.5 (Eq/ml)
105
104 103
102
10
1 1
10
102
103
104
105
106
bDNA (Eq/ml) • Monitor
o
Monitor 1.5
107
VIH-1 subtype B B B B B B B B B B B B A A A A A A A A A A A A A A A A A A D D E F G G
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