HLA and Drug Resistance Thomas Harrer Dept. of Medicine 3 - - PowerPoint PPT Presentation

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HLA and Drug Resistance Thomas Harrer Dept. of Medicine 3 - - PowerPoint PPT Presentation

Oct 02, 2022 136 likes •411 views

HLA and Drug Resistance Thomas Harrer Dept. of Medicine 3 University Hospital Erlangen Sandra Mller Birgit Schtz Bernd Spriewald Kathrin Eismann Hauke Walter Silke Bergmann Klaus Korn Ellen Harrer Barbara Schmidt Michael Buerle

slide-1
SLIDE 1

HLA and Drug Resistance

Thomas Harrer

  • Dept. of Medicine 3

University Hospital Erlangen Sandra Müller

Birgit Schätz Bernd Spriewald Kathrin Eismann Hauke Walter Silke Bergmann Klaus Korn Ellen Harrer Barbara Schmidt Michael Bäuerle Heinrich Sticht

SFB 466

slide-2
SLIDE 2

TCR

CTL

HIV

HLA A 2 HLA CW 8 HLA CW 4 HLA A3 HLA B7 HLA B27

slide-3
SLIDE 3

Gag Pol vif Env

Tat Rev Nef

vpr vpu

LTR LTR HLA A2,A3 B27,B8 Cw3,4 HLA A24,A32 B14,B51 Cw5,8

slide-4
SLIDE 4

I V Y Q Y V D D L I

TCR

CTL

  • IVY

Q Y V D D L I

TCR

CTL

+

Schmitt et al, AIDS 2000

V I I Y Q M Y D D L

TCR

CTL

+

Harrer E et al, JID 1996

Influence of M184V 3TC-Resistance mutation on CTL-recognition

slide-5
SLIDE 5
  • V

I I Y Q YM D D L

TCR

CTL

+

E I I Y QY M D D L

slide-6
SLIDE 6

L A V E I C T E L

TCR

CTL

+

  • V

I E C A M T E L

AZT-Resistance mutation M41L Gnerates an HLA-A2 Neo-Epitope: Wilde type: ALVEICTEM does not bind to HLA A2 mutated ALVEICTEL binds to HLA A2 Samri et al, JVi 2000

slide-7
SLIDE 7

Correlations between AA substitutions in the PR and HLA class I alleles in 94 patients

described epitopes potential epitopes major mutations minor mutations mutations are classified according to Johnson et al. 2005

♦mutations contributing to TPV or ATV

resistance relevant correlations

L19 1 5 L10 10 20 30 40 50 60 89% 88% 83% 10 I13♦ I15 15

PR variability in 94 patients (%)

20 25 K20 L24 30 40 35 P39 K43♦ 45 M46 I47 I54 55 50 60 I62♦ G73 I93♦ 65 70 V82 L90 I84 75 80 85 90 95

amino acids

Negative HLA class I associations Published and potential HLA class I restricted epitopes A2 B44 B62 B13 A23 A11 A2 A2 B44 B62 B62 A2 A74 A3/A11 A2/Cw6 B13 A2

D30 L33 I50 A71Δ V77Δ

A19 A68 A2 B51 A2 A3 B44 B62 A2

G48

B44 Cw2 B51 Cw5 A23 Cw3 A3

T74♦

Cw6 Cw3 Cw3 Cw4 Cw6 Cw3

  • A68

Positive HLA class I associations

V3 L63 I64

B62

M41

B44 Cw7

T12 N37

  • V32

K14 M36 F53 R70 N88 L89 G16♦ E35♦ Q58♦ D60♦ H69♦ N83♦

Cw3 B62 B18 A26 B44 Cw6

E65

A3/A11 A2 B44

Müller et al, JVI 2007

slide-8
SLIDE 8

Relevant correlations between HLA class I alleles and amino acid substitutions in the PR

position HLA P-value OR n* known epitopes potential epitopes** major mutations L33♦Δ A2 0.005 12.6 9 A2 M46♦Δ A2 0.049 2.9 14 +/A2 minor mutations L10♦Δ Cw2 0.032 4.0 9 A71Δ B44 0.013 3.5 14 B44 V77 B62 0.037 3.5 9 polymorphisms I13♦ A11 0.012 5.6 6 +/A3 supertype I13♦ B62 0.018 0.0027 I13♦ B13 0.020 8.7 4 B13 K14 B51 0.001 13.0 6 B51 I15 A23 0.012 7.5 5 I15 Cw7 0.019 0.27 6 Cw7 E35♦ B44 1.2x10E-9 34.4 21 +/B44 E35♦ Cw5 0.000014 2543.6 10 P39 B44 0.048 6.1 4 +/B44 P39 B18 0.046 10.6 2 B18 R41 A26 0.032 9.1 3 A26 K43♦ B62 0.012 6.7 5 B62 K43♦ Cw3 0.000073 21.5 8 Cw3 I54♦ B44 0.017 4.6 7 B44 I62Δ A2 0.027 3.0 20 A2 I62Δ A23 0.028 0.0017 I64 A3 0.030 0.2 2 A3 E65 Cw6 0.045 6.3 3 Cw6 T74♦ Cw6 0.028 8.4 3 Cw6 I93Δ B62 2.0x10E-6 31.5 15 B62 I93Δ Cw3 0.00038 8.9 17 Cw3

univariate statistical analyses (Fisher’s exact test) Bonferroni correction: p≤0.00003 biologically relevant correlations may be missed to confirm correlations biological assays were conducted

slide-9
SLIDE 9 S F U / 1 c e l s S F U / 1 c e l s S F U / 1 c e l s

EW9 DI EW9 E/D EW9 N/D EW9 M/I EW9

S F U / 1 c e l s

200 400 600 800

  • D-------
  • DI------
  • --D-----
  • -I------
  • ----S---
  • ------K-

no peptide EEMNLPGRW

SFU/100 000 cells

200 400 600 800

  • D-------
  • DI------
  • --D-----
  • -I------
  • ----S---
  • ------K-

no peptide EEMNLPGRW

SFU/100 000 cells

50 150 200 250

  • D-------
  • DI------
  • --D-----
  • -I------
  • ----S---
  • ------K-

no peptide EEMNLPGRW

100

SFU/100 000 cells

Recognition of EEMNLPGRW and variant peptides

slide-10
SLIDE 10

E35D: polymorphism

E35D negative effect on substrate binding affinity: Meiselbach et al. 2006 Reduced in vitro activity of ritonavir/amprenavir Associated with early treatment failure to PIs (Alexander et al. AIDS 2001)

slide-11
SLIDE 11

Recognition of newly predicted epitopes containing HLA class I linked mutations

Mutations correlating to HLA class I alleles are shown in red

aa position HLA m otif frequency of recognition

K14 B51 Q RPLVTVKIG 11/16 L33 A2 LLDTG ADDTVL 29/76 K43 Cw 3 LPG RW KPKM I 4/15 K43 Cw 3 W KPKM IG G I 6/12 I62 A2 KVRQ YDQ IL 25/76 I64 A3 ILIEICG HK * 17/33 I62 B13 RQ YDQ IPIEI* 12/18 E65 B13 RQ YDQ IPIEI* 12/18 K70 A3 ILIEICG HK * 17/33 A71 B44 IEICG HK AIG 12/23 I93 B62 TQ IG CTLNF 4/6 I93 Cw 3 TQ IG CTLNF 4/9 I93 B62/Cw 3 TQ IG CTLNF 16/17

slide-12
SLIDE 12

Einfluss von Mutationen auf die CTL Erkennung

w ild-type sequence m utant peptides H L A im pairm ent of C D 8+ T-cell activity* im provem ent

  • f C D 8+ T-cell

activity *

Q R P LV TV K IG Q R P LV TV R IG B 51 5/6 0/6 LLD TG AD D TV L LLD TG AD D TV F A2 6/12 2/12 LLD TG AD D TV L LID TG AD D TV F A2 4/12 7/12 K V R Q YD Q IL K V R Q YD Q V L A2 0/2 1/2 R Q YD Q IP IE I R Q YD Q V P IE I B 13 6/8 2/8 ILIE IC G H K ILIE IC G H R A3 9/12 1/12 IE IC G H K AIG IE IC G H K V IG B 44 3/11 7/11 TQ IG C TLN F TQ LG C TLN F B 62 3/6 3/6 TQ IG C TLN F TQ LG C TLN F B 62/ C w 3 5/12 4/12

Mutations correlating to HLA class I alleles are shown in red

slide-13
SLIDE 13

Recognition of KI10 and M46I / M54V mutants

KMIGGIGGFI (KI10) is highly conserved Contains the resistance mutations M46I/L and I54V M46I can abrogate CTL recognition = CTL escape mutation M46I in HLA A2+ Patienten more frequently (OR 2,9)

S F U / 1 . c e l s

500 1000 1500

KMIGGIGGFI no peptide

  • --------V
  • I--------
S F U / 1 . c e l s

no peptide KMIGGIGGFI

  • --------V
  • I--------

200 400 600 800

S F U / 1 . c e l s

200 400 600 800

KMIGGIGGFI

  • --------V
  • I--------

no peptide

S F U / 1 . c e l s

KMIGGIGGFI

  • I--------

200 400 600 800

no peptide

SFU/100 000 cells SFU/100 000 cells SFU/100 000 cells SFU/100 000 cells

KI10-I KI10

slide-14
SLIDE 14

Summary

  • Significant

Significant associati ssociation of frequent f frequent HLA alleles HLA alleles with occurrence ccurrence of defined f defined resis resistance ance mutations mutations and pol nd polymorphism morphisms

  • Definiti

Definition of ten new

  • n of ten new CTL epi

TL epitopes topes

  • Dru

Drug escap escape variants variants may may inhibit nhibit or

  • r au

augment gment the the CTL response, dependent CTL response, dependent

  • n t

the T e T-cell r l repertoire o e of t the p e patients.

  • Some

Some patients patients mount mount an oligoclonal n oligoclonal immune response mmune response targeting targeting dru rug resis resistant ant viruses iruses

  • CTL induce

CTL induce muta mutati tions

  • ns presumably

resumably have ave an important n important influence nfluence on the

  • n the

development development of drug

  • f drug mutati

mutation

  • ns and mutati

nd mutational

  • nal pathways

pathways

slide-15
SLIDE 15

Selection by ART and CTL

HIV-Genome ART ART + CTL CTL

slide-16
SLIDE 16

HIV HIV-

  • 1

1 Protease Protease: : Structure Structure and and Resistance Resistance

~ ¼ of all amino acids can mutate: Drug resistance and Polymorphisms

: Los Alamos. http://hiv-web.lanl.gov/

slide-17
SLIDE 17

Protease Inhibitor Resistance

90 77 73 54 46 36 32 24 20 10

IDV

L L K V M I A V I 1 IRV MR I I IL V VT SA I AFTS V M 99 82 71 M G V L I 84

APV

84 47 32 10 I V LMV 46 50 54 FIRV I V IL I V V FIRV 90 77 71 54 46 36 33 20

RTV

82 32 L F IL VL 84 10 MR I VT I AFTSV M 90 77 73 54 48 10

SQV

G 71 82 V S A 84 IRV M VL VT I V 77 71 46 36 30 10

NFV

L D FI N IL AFTS V 82 88 84 M I VT I 90 N DS 90 71 54 46 20

LPV/RTV

82 IL VL 84 10 MR L I 24 53 L F AFTS V M VT FIRV

Antiretroviral Resistance Mutations, A collaborative effort of the International AIDS Society-USA Resistance Testing Panel and HIV

  • InSite. HIV Clinical Trials 2001;2(4):346-355
slide-18
SLIDE 18

Influence of resistance associated mutations on CD8+ T-cell recognition

no peptide

  • -----V----

VRQYDQIPIEI

100 200 300 400 500 600

SFU/100 000 cells

VI11-V VI11

no peptide

  • ------V--

IEICGHKAIG

IG10-V IG10

SFU/100 000 cells

50 100 150 200 200 400 600 800

no peptide.

  • -----V----

VRQYDQIPIEI SFU/100 000 cells

VI11-V VI11

S F U / 1 c e l s S F U / 1 c e l s

no peptide

100 200 300 400 500 600

  • -L------

TQIGCTLNF SFU/100 000 cells

TF9-L TF9

slide-19
SLIDE 19

Priming of M46I-Viruses by CTL

strong CTL-response against KI10: Selection of M46I because of strong fitness disadvantage wild type M46 more prevalent in plasma

  • n therapy wild type decreases and M46I prevails

hypothesis: HLA A2+ u. B62 + patients develop M46I – Mutation more rapidly planned: Prospective study in patients failing PIs with M46I

slide-20
SLIDE 20

Summary

  • resistance mutations/polymorphisms correlating with

HLA-types acted as escape mutations in most patients

  • several patients showed recognition of variant peptides probably due

to generation of new TCR-specificities

  • in some patients antigenicity was sustained or even enhanced

despite of mutations in the epitopes

univariate statistical analyses with “high” p-values can indicate T-cell epitopes

  • 25 relevant (p<0.05) correlations between HLA class I

alleles und the occurrence of drug resistance mutations/polymorphisms in PR

  • all tested epitopes were confirmed in biological assays

biologically relevant associations are missed due to Bonferroni correction

slide-21
SLIDE 21

Resistance mutations

M D K LT K 41 67 70 210 215 219 N R W YF QE L K L M 65 74 184 65 69 74 184 44 118 184 75 74 6567 70 115 184 210 215 219 D I TMSA F V E V D T R V Y V I V

AZT ddI ddC 3TC d4T ABC

M D K LT K 41 67 70 210 215 219 N R W YF QE L L R N R V V R W YF QE V V

Antiretroviral Resistance Mutations, A collaborative effort of the International AIDS Society-USA Resistance Testing Panel and HIV

  • InSite. HIV Clinical Trials 2001;2(4):346-355

K R 65

TDF

slide-22
SLIDE 22

Factors influencing the shaping of viral sequence by CTL

  • Epitope:
  • highly conserved, essential sequence
  • no escape mutations, viral control
  • conserved functional important sequence
  • mutations only by strong CTL selection
  • highly variable polymorphic regions
  • random mutations > escape mutations
slide-23
SLIDE 23

Factors influencing the shaping of viral sequence by CTL

Selection pressure by CTL

  • frequency of CTL
  • breadth of response: polyclonal ?
  • TCR avidity
  • TCR cross-reactivity
  • functional CTL resistance of HIV-1
slide-24
SLIDE 24

Specific Recognition of the important K103N mutation

A*2301, A*3301, B*1501 and B*5801, A*2301 can be predicted to have the highest binding affinity for KKNKSVTVL

slide-25
SLIDE 25

CTL

HLA- I

infected Monocyte

TCR

HIV

  • lysis of infected cells

Granzyme, Fas-Ligand

  • secretion of cytokines
  • secretion of antiviral molecules

Rantes, MIP-1alpha

slide-26
SLIDE 26

Recognition of EEMNLPGRW and variant peptides

S F U / 1 c e l s S F U / 1 c e l s S F U / 1 c e l s S F U / 1 c e l s

500 1000 1500 2000

  • D-------
  • DI------
  • --D-----
  • -I------
  • ----S---
  • ------K-

no peptide EEMNLPGRW

SFU/100 000 cells

100 200 300 400 500

  • D-------
  • DI------
  • --D-----
  • -I------
  • ----S---
  • ------K-

no peptide EEMNLPGRW

SFU/100 000 cells

500 1000 1500 2000 2500 3000

  • D-------
  • DI------
  • --D-----
  • -I------
  • ----S---
  • ------K-

no peptide EEMNLPGRW

SFU/100 000 cells