1 Bcell responses to 13 valent pneumococcal conjugate

1 B-cell responses to 13 -valent pneumococcal conjugate vaccine in patients with severe chronic kidney disease Gabrielle Gaultier, Ph. D Biotechnology Candidate Lakehead University Supervisor: Dr. Marina Ulanova Northern Health Research Conference 2017

Faculty/ Presenter Disclosure Slide Gabrielle Gaultier • Relationships with commercial interests: NONE • Potential for conflict(s) of interest: NONE 2

Pneumovax and Prevnar 13 3 • In Canada for all adults at risk for invasive pneumococcal disease including CKD patients, 1 dose of Pneumovax is recommended • Prevnar 13 is recommended for children and is part of the routine infant immunization schedule • Prevnar 13 is approved for adults, only recommended for specific groups (ex. HIV positive adults) Pneumovax (PPV 23) Prevnar 13 (PCV 13) Unconjugated pneumococcal polysaccharide vaccine Pneumococcal polysaccharide-protein conjugated vaccine (Diphtheria CRM 197 protein) 23 serotypes 13 serotypes (1, 2, 3, 4, 5, 6 B, 7 F, 8, 9 N, 9 V, 10 A, 11 A, 12 F, 14, (3, 4, 5, 6 A, 6 B, 7 F, 9 V, 14, 18 C, 19 A, 19 F and 15 B, 17 F, 18 C, 19 A, 19 F, 20, 22 F, 23 F, 33 F) 23 F) T cell independent response T cell dependent response http: //www. phac-aspc. gc. ca/publicat/cig-gci/p 04 -pneu-eng. php

Problems with Pneumovax 4 Katz, L. A. , et al. (1980), Nikoskelainen, Jukka, et al. (1985), Rytel, Michael W. , et al. (1986) • Responsiveness to vaccination in patients with CKD can be diminished • Pneumovax, is less effective in CKD patients than in healthy adults • Lower post-vaccination antibody levels, which rapidly decline within 6 months of vaccination Clutterbuck E. A. et al. (2012) • Immunization with Pneumovax in elderly, decrease in memory B-cell frequency • Memory B-cell responses to subsequent immunization with PCV 7, after Pneumovax were attenuated • Antibody hyporesponsiveness can be due to the depletion of the peripheral memory Bcell pool http: //www. sesml. org. au/latest-news/pneumovax-23 -pre-filled-syringe-presentation-20 -october-2015

5 Research Question Does previous immunization with PPV 23 affect the production of memory B cells after immunization with PCV 13 in adult patients with CKD? Objectives 1. Compare numbers of circulating memory B cells between PPV 23 naïve and those previously immunized with PPV 23 > 1 year ago.

Methods 6 Groups (n=30 per group) 1. CKD & PPV 23 naïve 2. CKD & previously immunized with PPV 23 > 1 year Experiment Outline Flow Cytometry Channel Measurement Forward-scattered light (FSC) Cell size Side-scattered light (SSC) Cell granularity FL 1 (FITC, 519 nm) CD 5 FL 2 (PE, 578 nm) CD 19 FL 3 (Per. CP/ Cy 5. 5, 695 nm) CD 27 FL 4 (APC, 785 nm) Ig. M

Results Addressing Objective: 7 Compare numbers of circulating B memory cells between PPV 23 naïve and those previously immunized with PPV 23 > 1 year ago. Group N Age Sex PPV 23 naïve 32 Mean= 64 Range: 33 -88 M= 19 F= 13 Immunized with PPV 23 > 1 year ago 24 Mean= 61 Range: 33 -89 M= 11 F= 13 Statistical analysis was performed with Graph. Pad Prism using the Mann-Whitney U test, no significant differences found.

Proportions of B cell Subpopulations 8 Proportions of B cell subpopulations 7 days post immunization Proportions of B cell subpopulations pre immunization Naïve B Cells 23% 27% Ig. M Memory B cells 52% 16% 49% Class Switched B cells Class Switched Memory B cells 8% 15% 9% Type of B cell Description Naïve (CD 19+CD 27 -Ig. M+) B cell that has never encountered an antigen Ig. M Memory (CD 19+CD 27+Ig. M+) Short lived B cell resulting from a T-independent response Class Switched (CD 19+CD 27 -Ig. M-) Activated B cell, naïve B cell that has recently encountered an antigen Class Switched Memory (CD 19+CD 27+Ig. M-) Long lived B cell resulting from a T-dependent response, responds rapidly upon re-exposure to antigen

Absolute Numbers of B cell Subpopulations Statistical analysis was performed with Graph. Pad Prism using the Mann-Whitney U test, no significant differences found. 9

CD 19+ CD 5+/- B cells CD 19+ CD 5 - cells have an important role in the Ig. G antibody response to pneumococcal capsular polysaccharides (Moens L. et al. 2015). Type of B cell Description B-1 a (CD 19+CD 5+) Responsible for secretion of Ig. M antibodies B-1 b (CD 19+CD 5 -) Responsible for secretion of Ig. M, Ig. A and Ig. G antibodies after antigen specific stimulation Statistical analysis was performed with Graph. Pad Prism using the Mann-Whitney U test, no significant differences found. 10

Conclusions 11 1. In adult CKD patients immunized with PCV 13, there is no difference in B cell subpopulations or total B cell numbers between those who received PPV 23 over one year ago and pneumococcal vaccine naïve participants. 2. These data suggest that previous immunization with PPV 23 should not affect the abilities of adult CKD patients to respond to immunization with PCV 13. 3. The groups are uneven because of lack of patients previously immunized with PPV 23. Although PPV 23 is the standard of care for CKD patients, only 45/127 enrolled in the study have been previously immunized.

12 Current NACI recommendations for immunization of adults against pneumococcal infections NACI October 2016

Acknowledgements • Dr. Marina Ulanova • Dr. William Mc. Cready • Dr. Eli Nix • Angele Desbiens-Forget • Clinical trials nurses and Hemodialysis nurses Funding 13