Chakrabarti, B

Douagi, I

Forsell, MN

Hedestam, GB Karlsson

Loré, K

Soldemo, M

Sundling, C

Wyatt, RT

English

PubMed

Douagi I

Wyatt RT

Loré K

Forsell MN

Chakrabarti B

Soldemo M

Sundling C

Hedestam GB

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Retrovirology

P05-07. Evaluation of peripheral and bone marrow B cell responses in rhesus macaques after immunization with soluble HIV-1 gp140 trimers

C Sundling

M Soldemo

B Chakrabarti

MN Forsell

K Loré

RT Wyatt

I Douagi

GB Karlsson Hedestam

Crossref

BioMed Central
Page 1 of 1
(page number not for citation purposes)
Retrovirology
Open AccessPoster presentation
P05-07. Evaluation of peripheral and bone marrow B cell responses 
in rhesus macaques after immunization with soluble HIV-1 gp140 
trimers
C Sundling*1, M Soldemo1, B Chakrabarti2, MN Forsell2, K Loré3, RT Wyatt2, 
I Douagi1 and GB Karlsson Hedestam1
Address: 1Microbiology, Tumor and Cell Biology, Karolinska Institutet, Stockholm, Sweden, 2Vaccine Research Center, National Institutes of 
Health, Bethesda, MD, USA and 3Center for Infectious Medicine, Karolinska Institutet, Stockholm, Sweden
* Corresponding author    
Background
Broadly neutralizing antibodies (bNAbs) against HIV-1
are not efficiently elicited by vaccination despite consider-
able efforts. bNAb elicitation is likely limited both by cur-
rent envelope glycoproteins (Env) immunogens, which
are not sufficient mimics of the native Env spike complex,
and by the lack of immunization regimens that promote
optimal development and maturation of B cell responses.
So far, analyses from protein immunization studies focus
mainly on serological antibody responses and less on
responses at the B cell level. Here, we characterize frequen-
cies of antigen-specific antibody secreting cells (ASC) and
memory B cells in the periphery and bone marrow of rhe-
sus macaques immunized with soluble gp140 trimers.
These analyses provide a foundation for the design of
improved immunization protocols to elicit bNAbs against
HIV-1.
Methods
We established stimulation protocols for in vitro expan-
sion and differentiation of macaque memory B cells into
ASC to allow enumeration and monitoring of memory B
cells post immunization. Using a series of variant Env
probes, the frequencies of B cells specific for different
structural determinants of Env were quantitated by B cell
ELISpot analysis.
Results
By stimulating peripheral PBMCs with a mitogen cocktail
we detected Env-specific memory B cells at a sensitivity of
~0.1% of the total number of memory cells. By sampling
peripheral PBMCs at several time points after soluble
gp140 trimer immunization we demonstrate that ASC
peak one week after immunization while memory B cell
responses peak between one and two weeks after immuni-
zation. The peripheral gp140-specific memory B cell pool
constitutes between 10 to 20% of the total peripheral
memory B cell pool.
Conclusion
By establishing in vitro stimulation protocols for rhesus
memory B cells, and determining the peak of antigen spe-
cific responses after protein vaccination, we describe a
benchmark study that can be used to inform future analy-
ses of vaccine-elicited B cell responses in primates.
from AIDS Vaccine 2009
Paris, France. 19–22 October 2009
Published: 22 October 2009
Retrovirology 2009, 6(Suppl 3):P83 doi:10.1186/1742-4690-6-S3-P83
<supplement> <title> <p>AIDS Vaccine 2009</p> </title> <editor>Anna Laura Ross</editor> <note>Meeting abstracts – A single PDF containing all abstracts in this Supplement is available <a href="http://www.biomedcentral.com/content/files/pdf/1742-4690-6-S3-full.pdf">here</a>.</note> <url>http://www.biomedcentral.com/content/pdf/1471-2105-10-S12-info.pdf</url> </supplement>
This abstract is available from: http://www.retrovirology.com/content/6/S3/P83
© 2009 Sundling et al; licensee BioMed Central Ltd. 


GB Hedestam

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P05-07. Evaluation of peripheral and bone marrow B cell responses in rhesus macaques after immunization with soluble HIV-1 gp140 trimers

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