Humoral and lung immune responses to Mycobacterium tuberculosis infection in a primate model of protection q,qq Noton K. Dutta a,1,2 , James McLachlan b,1 , Smriti Mehra a,b , Deepak Kaushal a,b,⇑ a Tulane National Primate Research Center, Covington, LA, USA b Department of Microbiology & Immunology, Tulane University Health Sciences Center, New Orleans, LA, USA a r t i c l e i n f o Article history: Received 8 November 2013 Revised 11 February 2014 Accepted 17 February 2014 Keywords: M. tuberculosis BCG Animal model Granuloma Macrophage Lymphocyte a b s t r a c t Rece ntly we repo rted (Mehra et al., 2013), tha t lung granu lomas from Mycobacterium bovis Bacille Calme tte–Guér in (BCG)-v accinated cynomolgus macaques exhibit upon challenge with M. tuberculosis a more bala nce d exp ression ofa- an d b-che mok ines , relative to compara ble samples from sham - vaccinated animals by comparative transcriptomics. Here, we studied the recruitment of immune cells to bl oo d an d lung s in M. tube rculos is-inf ecte d mac aqu es as a function of prio r BCG-vaccination . Vaccination initially enhanced the levels of both macrophages and lymphocytes in blood. In contrast, significan tly more CD4 + lympho cyte s wer e later recruited to the lun gs of sham-vaccinated anim als compared with earlier times/BCG vaccinated animals. BCG-vaccination had a short-lived impact on the anti-M. tuberculosis response. M. tuberculosis cont inu ed to repl icate in the lun g even in the wake ofincreased CD4 + T cell recruitment to primate lungs, indicating that immune subversive mechanis ms are key to its survival in vivo. 2014 The Aut hors . Publ ishe d by Else vier Ltd. Thi s is anopen acce ss arti cleundertheCC BY-NC- NDlicens e (http://creativecommons.org/licenses/by-nc-nd/3.0/). Introduction Tub erculosis is responsible for the death of over 1.5 mil lion peo ple eve ry year [1] . Thi s situ atio n has worse ned due to the emerg ence of dru g-resistanc e [2] , HI V co-i nf ection an d th e insufficient pro tect ion of the Bacillus Calmette –G uér in (BC G) vaccine, which is unable to protect adults against pulmonary TB [3,4] . The efficac y of BCG in pro tecting against adu lt pul mon ary TB is unsatisfactory [5] . New and efficacious vaccines against TB are therefore urgen tly needed [6,7]. Thi s req uire s both a better und erst and ing of the corr elat es of pro tect ion in app rop riat e experim ental model s [8,9], and also a clearer understanding ofth e shor tco mi ng s of BC G such that th ey can be av oid ed wi th fut ur e vaccine design. Nonhuman Primates (NHP s), such as rhesus[10–13] or cyno- molgus [11,14] macaques are excellent models ofMycobacterium tuberculosis(Mtb) infection [15]. Upon experimental infection via either the intrabr onc hia l or the aero sol rou te, these anim als re capitu lat e the entire br ead th of the hu ma n TB infec tio n syn dro me , includi ng acute TB [10] chara cter ize d by massive caseous immu nopat holog y [13] and asym pto mat ic, late nt infe ctio n (LTBI). Furthermore, in macaques, LTBI can be reactivated by SIV co-infection [12], TNFa inhibition [12,16] or CD4-d epletion[17]. More imp orta ntl y, macaques dem ons trat e a spectrum of lung pat hol ogi cal out comes upo n exp erim ent al infe ction wit h Mtb, similar to naturally infect ed human beings[10–13,15,18–20] . Akin to humans, vaccination with BCG can protect NHPs against Mtb challen ge, but the pro tect ion is inco mp lete [11,21,22]. Th ese ob ser - vations reinforce our contention that macaques best represent the human TB infection syndrome, allowing a study of pathology as well as protective responses not possible in other model systems [23]. It is known that prior BCG vaccination leads to a higher expres- sion ofb-chemokines in lung lesions followin g Mtb challenge[11]. http://dx.doi.org/10.1016/j.trivac.2014.02.001 1879-4378/2014 The Authors. Published by Elsevier Ltd. Thi s is an ope n acce ss article un der the CC BY- NC- ND lice nse (http://creativecommons.org/licenses/by-nc-nd/3.0/ ). Abbreviations:NHP, Nonhuman Primate; TB, tuberculosis; Mtb, Mycobacterium tuberculosis ; BCG, Bacille Calmette–Guerin; BAL, broncho-alveolar lavage. q The Tulane Nation al Primate Resea rch Center facili ties are accre dited by the American Association for Accreditation of Laboratory Animal Care and licensed by the US Depa rtme nt of Agricultu re. All animals were rout inel y cared for accord ing to the guidelines prescribed by the NIH Guide to Laboratory Animal Care. NHP studies were conducted following the recommendatio ns of the institutional animal care and use committee. Humane endpoints were pre-defined in this protocol. qq The procedu res repo rted in this manus cript were appro ved by the relevant oversight committees (IACUC and IBC). ⇑ Corresponding author at: Department of Microbiology & Immunology, Tulane National Primate Research Center, 18703 Three Rivers Road, Covington, LA 70433, USA. Tel.: +1 (985) 871 6254; fax: +1 (985) 871 6260. E-mail address: [email protected](D. Kaushal). 1 Contributed equally. 2 Pres ent addre ss: Cent er for Tub erculo sis Research, Johns Hopk ins School ofMedicine, Baltimore, MD, USA. Trials in Vaccinology 3 (2014) 47–51 Contents lists available at ScienceDirect Trials in Vaccinology journal homepage: www.elsevier.com/locate/trivac
5
Embed
Humoral and Lung Immune Responses to Mycobacterium Tuberculosis
This document is posted to help you gain knowledge. Please leave a comment to let me know what you think about it! Share it to your friends and learn new things together.
Transcript
8/10/2019 Humoral and Lung Immune Responses to Mycobacterium Tuberculosis
Humoral and lung immune responses to Mycobacterium tuberculosis
infection in a primate model of protectionq,qq
Noton K. Dutta a,1,2, James McLachlan b,1, Smriti Mehra a,b, Deepak Kaushal a,b,⇑
a Tulane National Primate Research Center, Covington, LA, USAb Department of Microbiology & Immunology, Tulane University Health Sciences Center, New Orleans, LA, USA
a r t i c l e i n f o
Article history:
Received 8 November 2013
Revised 11 February 2014
Accepted 17 February 2014
Keywords:
M. tuberculosis
BCG
Animal model
Granuloma
Macrophage
Lymphocyte
a b s t r a c t
Recently we reported (Mehra et al., 2013), that lung granulomas from Mycobacterium bovis Bacille
Calmette–Guérin (BCG)-vaccinated cynomolgus macaques exhibit upon challenge with M. tuberculosis
a more balanced expression of a- and b-chemokines, relative to comparable samples from sham-
vaccinated animals by comparative transcriptomics. Here, we studied the recruitment of immune cells
to blood and lungs in M. tuberculosis-infected macaques as a function of prior BCG-vaccination.
Vaccination initially enhanced the levels of both macrophages and lymphocytes in blood. In contrast,
significantly more CD4+ lymphocytes were later recruited to the lungs of sham-vaccinated animals
compared with earlier times/BCG vaccinated animals. BCG-vaccination had a short-lived impact on the
anti-M. tuberculosis response. M. tuberculosis continued to replicate in the lung even in the wake of
increased CD4+ T cell recruitment to primate lungs, indicating that immune subversive mechanisms
are key to its survival in vivo.
2014 The Authors. Published by Elsevier Ltd. This is anopen access articleundertheCC BY-NC-NDlicense
Tuberculosis is responsible for the death of over 1.5 million
people every year [1]. This situation has worsened due to the
emergence of drug-resistance [2], HIV co-infection and the
insufficient protection of the Bacillus Calmette–Guérin (BCG)
vaccine, which is unable to protect adults against pulmonary TB
[3,4]. The efficacy of BCG in protecting against adult pulmonary
TB is unsatisfactory [5]. New and efficacious vaccines against TB
are therefore urgently needed [6,7]. This requires both a better
understanding of the correlates of protection in appropriate
experimental models [8,9], and also a clearer understanding of
the shortcomings of BCG such that they can be avoided with future
vaccine design.
Nonhuman Primates (NHPs), such as rhesus [10–13] or cyno-
molgus [11,14] macaques are excellent models of Mycobacterium
tuberculosis (Mtb) infection [15]. Upon experimental infection via
either the intrabronchial or the aerosol route, these animals
recapitulate the entire breadth of the human TB infection
syndrome, including acute TB [10] characterized by massive
caseous immunopathology [13] and asymptomatic, latent infection
(LTBI). Furthermore, in macaques, LTBI can be reactivated by SIVco-infection [12], TNFa inhibition [12,16] or CD4-depletion [17].
More importantly, macaques demonstrate a spectrum of lung
pathological outcomes upon experimental infection with Mtb,
similar to naturally infected human beings [10–13,15,18–20]. Akin
to humans, vaccination with BCG can protect NHPs against Mtb
challenge, but the protection is incomplete [11,21,22]. These obser-
vations reinforce our contention that macaques best represent the
human TB infection syndrome, allowing a study of pathology as
well as protective responses not possible in other model systems
[23].
It is known that prior BCG vaccination leads to a higher expres-
sion of b-chemokines in lung lesions following Mtb challenge [11].
http://dx.doi.org/10.1016/j.trivac.2014.02.001
1879-4378/ 2014 The Authors. Published by Elsevier Ltd.This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/3.0/ ).
q The Tulane National Primate Research Center facilities are accredited by theAmerican Association for Accreditation of Laboratory Animal Care and licensed by
the US Department of Agriculture. All animals were routinely cared for according to
the guidelines prescribed by the NIH Guide to Laboratory Animal Care. NHP studies
were conducted following the recommendations of the institutional animal care
and use committee. Humane endpoints were pre-defined in this protocol.qq The procedures reported in this manuscript were approved by the relevant
oversight committees (IACUC and IBC).⇑ Corresponding author at: Department of Microbiology & Immunology, Tulane
National Primate Research Center, 18703 Three Rivers Road, Covington, LA 70433,
curred in sham-vaccinated animals, in an antigen load-dependent
manner. Moreover, immunophenotyping of both lung tissue and
BAL showed higher CD4+ T cell counts in sham-vaccinated relative
to BCG-vaccinated macaques during early time-points (Fig. 2). The
differences manifested themselves primarily in the granulomatouslesions obtained from sham-vaccinated relative to BCG-vaccinated
macaques at week 5 and further increased at week 10. The differ-
ences were not observed for the surrounding non-granulomatous
normal tissue and were specific to lesion tissue only. Significant dif-
ferences were also not observed for CD8+ T cells at either 5 or
10 weeks. Thus, recruitment and accumulation of likely antigen-
specific, Th1-type, activated CD4+ T cells in granulomatous lesions
occurred as a function of antigen load. However, the accumulation
of these CD4+ T cells in the granulomas of sham-vaccinated NHPs
failed to check both the replication of Mtb and the onset of acute
TB at this site [11].
Since CD4+ T cells are absolutely required for the protective im-
mune response to Mtb infection [34], our results suggest that their
recruitment to the lung by itself is insufficient in controlling theinfection. This underscores the ability of Mtb in modulating host
Fig. 4. CD4+ T cells expressedas a % of total lymphocytes areshown forlung (granulomaas well as non-granuloma) samples onthe Y -axis for BCG-vaccinated (red)and sham-
vaccinated (blue) NHPs at both 5 (A) and 10-week (B) post-Mtb infection. CD8+ T cells expressed as a % of total lymphocytes are shown for lung (granuloma as well as non-
granuloma) on the Y -axis for BCG-vaccinated(red)and sham-vaccinated (blue) NHPs at both 5 (C)and 10-week (D)post-Mtb infection. (For interpretation of the references to
color in this figure legend, the reader is referred to the web version of this article.)
50 N.K. Dutta et al./ Trials in Vaccinology 3 (2014) 47–51
8/10/2019 Humoral and Lung Immune Responses to Mycobacterium Tuberculosis
[21] M.H. Larsen, K. Biermann, B. Chen, T. Hsu, V.K. Sambandamurthy, A.A. Lackner,
et al., Efficacy and safety of live attenuated persistent and rapidly cleared
Mycobacterium tuberculosis vaccine candidates in non-human primates,
Vaccine 27 (2009) 4709–4717.
[22] F.A. Verreck, R.A. Vervenne, I. Kondova, K.W. van Kralingen, E.J. Remarque, G.
Braskamp, et al., MVA.85A boosting of BCG and an attenuated, phoP deficient
M. tuberculosis vaccine both show protective efficacy against tuberculosis in
rhesus macaques, PLoS ONE 4 (2009) e5264.[23] D. Kaushal, R.P. Bohm Jr., A.A. Lackner, How well do you knowyour monkeys?,
J Med. Primatol. 42 (2013) 48–49.
[24] S.H. Kaufmann, Tuberculosis vaccines: time to think about the nextgeneration,
Semin. Immunol. 25 (2013) 172–181.
[25] E.J. Rubin, The granuloma in tuberculosis – friend or foe?, N Engl. J. Med. 360
(2009) 2471–2473.
[26] L. Ramakrishnan, Revisitingthe role of the granuloma in tuberculosis, Nat. Rev.
Immunol. 12 (2012) 352–366.
[27] C. Paige, W.R. Bishai, Penitentiary or penthouse condo: the tuberculous
granuloma fromthe microbe’s point ofview, Cell.Microbiol. 12(2010)301–309.
[28] J.T. Mattila, O.O. Ojo, D. Kepka-Lenhart, S. Marino, J.H. Kim, S.Y. Eum, et al.,
Microenvironments in tuberculous granulomas are delineated by distinct
populations of macrophage subsets and expression of nitric oxide synthase
and arginase isoforms, J. Immunol. 191 (2013) 773–784.
[29] D.G. Russell, The evolutionary pressures that have molded Mycobacteriumtuberculosis into an infectious adjuvant, Curr. Opin. Microbiol. 16 (2013) 78–
84.
[30] D.G. Russell, P.J. Cardona, M.J. Kim, S. Allain, F. Altare, Foamy macrophages and
the progression of the human tuberculosis granuloma, Nat. Immunol. 10(2009) 943–948.
[31] J.M. Davis, L. Ramakrishnan, The role of the granuloma in expansion and
dissemination of early tuberculous infection, Cell 136 (2009) 37–49.
[32] R.F. Silver, J. Walrath, H. Lee, B.A. Jacobson, H. Horton, M.R. Bowman, et al.,
Human alveolar macrophage gene responses to Mycobacterium tuberculosisstrains H37Ra and H37Rv, Am. J. Respir. Cell Mol. Biol. 40 (2009) 491–504.
[33] M.A. Baird, D.N. Hart, N. Abernethy, J.D. Watson, Dendritic cell presentation of
PPD and19 kDa protein of Mycobacteriumtuberculosis and emergentT helper