Table 3.
Animal studies of probiotic therapies: summary of study characteristics and major findings
| Study | ROB | Sample | MS Model | Intervention | Timeline | Duration | Major Findings |
|---|---|---|---|---|---|---|---|
| Maassen et al.59 | UN | F SJL mice 8–12 wko n = 6/g |
PLP-induced EAE |
T1: L. reuteri ML1 T2: L. casei 393 T3: L. plantarum NCIB 8826 T4: L. murinus CNRZ Each 1010 CFU o.g. C: NaCO3 |
Proph | Every other day for 5 admins |
CD: ↑ CDB (T1), weak ↓ (T2), ↓ (T4), – (T3) IM: NA MM: NA MC: NA |
| Salehipour et al.50 | Low | F C57BL6 mice 8–10 wko n = 8/g |
MOG-induced EAE |
T1: L. plantarum A7, 109 CFU o.g. T2: B. animalis PTCC 1631,109 CFU o.g. T3: T1+ T2 C: sterile saline |
Thera | Daily for 22 d |
CD: delayed EAE onset (T1-T3), T3 more pronounced; ↓ EAE CS, CDI, incidence, infiltration of MNCs, and demyelination (T1-T3), T3 more pronounced IM: ↓ ASP, IFN-γ, IL-6, IL-17 (T1-T3), T3 more pronounced; ↑ % CD4+ CD25+ Foxp3+ Tregs, IL-4, IL-10, TGF-β (T1-T3), T3 more pronounced MM: NA MC: ↑ expression of GATA3, FoxP3 and ↓ expression of Tbet, RORγt (T1-T3), T3 more pronounced |
| He et al.75 | UN | F C57BL6 mice 10–12 wko n = 37–40/g |
MOG-induced EAE |
T: L. reuteri DSM 17938 100 µL of 108 CFU o.g. C1: 100 µL MRS media + EAE, o.g. C2: normal control |
Thera | Daily for 20 d |
CD: ↓ CDS, EAE incidence, maximum CS, inflammatory cell infiltration IM: ↓ CD3 + T cells & CD68+ macrophages in spinal cord, % and abs. # of TH1 and TH17 cells, IL-17, IFN-γ, ↓ % MOG35-55-specific splenocytes MM: reversed EAE rel. abund. changes (↑ Bacteriodetes, Proteobacteria, Deferribacteres); Bifidobacterium, Lactobacillus, Prevotella, & S24-7 = negative correlation w/ CS MC: NA |
| Goudarzvand et al.52 | UN | M Wistar rats 8–10 wko n = 8/g |
GID |
T1: L. plantarum T2: Bifidobacterium B94 Each 1.5 × 108 CFU/mL orally C1: GID only C2: sterile saline, no GID |
Thera | Daily for 28 d |
CD: – traveled distance, escape latency, or swimming speed IM: NA MM: NA MC: NA |
| Consonni et al.53 | Low | F Lewis rats 6–8 wko n = 6–9/g (T1-T4), n = 18/g (T5-T6) |
gpMBP-induced EAE |
T1: L. crispatus LMG P-23257 T2: L. rhamnosus ATCC 53103 T3: B. animalis subsp. lactis BB12 T4: B. animalis subsp. LMG S-28195 T5: T1+ T2 T6: T3+ T4 Each 2 × 109 CFU/300 µL orally C: vehicle |
Proph & Thera | 15 doses over 3 wks (5 doses pre-EAE) |
CD: ↓ EAE incidence & median score at peak (T1-T6); ↓ myelin loss, astrocytosis, & spinal cord immune cell infiltration (T5/T6); delayed EAE onset (T1,T2, T5, T6); dose-response relationship observed with T5/T6 IM: ↓ proliferative response to MBP, IFN-γ, TNF-α, IL-17 (T5/T6); ↑ TGF-β, IL-16 (T5/T6) MM: NA MC: NA |
| Baken et al.73 | UN | M Lewis rats 6–8 wko n = 8/g |
gpSCH-induced EAE |
T: L. casei strain Shirota, 1 mL of 1 × 109 CFU/mL C: 1 mL saline/peptone |
Proph & Thera | Daily for 35 d (starting 8 d pre-EAE) |
CD: ↓ body weight; ↑ EAE incidence, duration, CDS, & CDI; earlier EAE onset IM: NA MM: NA MC: NA |
| Gharehkhani Digehsara et al.71 | UN | F C57BL6 mice 8–10 wko |
Cuprizone (4 wks) |
T1: L. casei 4 wks, cuprizone 4 wks T2: Cuprizone 4 wks, L. casei 4 wks T3: Cuprizone 4 wks, L. casei 4 wks w/ Vitamin D3 (20 IU/d) C1: L. casei 4 wks, 1 × 109 CFU/mL C2: Cuprizone 4 wks, 0.2% w/w All admin orally C3: normal control |
Proph & Thera |
CD: more normal & significant Y-maze alternation behavior (T1-T3) IM: ↓ IL-17 (T1-T3), T3 more pronounced; ↑ TGF-β (T1-T3) MM: NA MC: ↓ expression of IDO gene & miR-155 (T1-T3, C2); ↑ expression of miR-25 (C2), trending in T1-T3 |
|
| Kobayashi et al.68 | UN | M & F Lewis rats 7 wko & 2 wko n = 8/g |
gpSCH- or gpMBP-induced EAE |
T1: L. casei strain Shirota in M rats (7 wko), 1–2 × 109 CFU o.g. T2: L. casei strain Shirota in F rats (7 wko), 1–2 × 109 CFU o.g. T3: L. casei strain Shirota in M & F rats (2 wko), 9.2–10.1 × 109 CFU o.g. T4: B. breve strain Yakult in M & F rats (2 wko), 5.0–6.9 × 109 CFU o.g. C: 0.5 mL saline/peptone |
Proph & Thera | Daily T1-T2: 35 d (starting 7 d pre-EAE) T3-T4: 63 d (starting 5 wks pre-EAE) |
CD: ↓ mortality in T1-T4 except T4 males (↑); – EAE onset, peak, mean CDS, infiltration of MNCs IM: – MBP IgG MM: NA MC: NA |
| Kobayashi et al.74 | UN | F SJL & C57BL6 mice 7 wko n = 15/g |
PLP-induced EAE (SJL) & MOG-induced EAE (C57BL6) |
T: L. casei YIT 9029, 0.6–1.2 × 109 CFU o.g. C: 0.2 mL saline/peptone |
Proph & Thera | Daily for 50 (SJL) or 29 (C57BL6) d (both starting 1 wk pre-EAE) |
CD: ↓ EAE CS on ds 12, 29, & 30 (SJL); – EAE onset, peak score, MNC infiltration, white matter demyelination, or neutrophil infiltration (SJL & C57BL6) IM: ↓ % CD8 + T cells in spleen (SJL), ↑ IL-10, % Tregs in spleen, IL-17, IFN-γ (SJL) MM: NA MC: NA |
| Lavasani et al.48 | UN | F C57BL6 (WT & IL-10-/-) mice 8–10 wko n = 3–18/g |
MOG-induced EAE |
T1: Proph L. paracasei DSM 13434 T2: Proph L. plantarum DSM 15312 T3: Proph L. plantarum DSM 15313 T4: Proph L. paracasei PCC 101 T5: Proph L. delbrueckii subsp. bulgaricus DSM 20081 Each 5 mL of 109 CFU orally until EAE, then 200 µL of 109 CFU o.g. T6: Thera T1 T7: Thera Lacto-mix (T1-T3) T8: HK T7 T9: T7 in IL-10-/- mice Each 200 µL of 109 CFU o.g. C: saline |
Proph & Thera | Daily for 37 d (starting 12 d pre-EAE) or every other day for 20 d (starting 2 wks post-EAE onset) |
CD: – EAE progression (T1-T5,T8); delayed EAE onset and ↓ CS (T1-T3, T7); therapeutic effects of Lacto-mix absent in T9 IM: ↓ T cell proliferation (T1-T3) ↓ CD4 + T cells (T1,T3),, ↓ IFN-γ, TNF-α (T1); ↑ IL-4, IL-10, TGF-β (T1); ↓ CNS inflammation, CD4 + T cell infiltration, IFN-γ, TNF-α, IL-17, and IL-17-producing CD4 + T cells (T7); ↑ IL-10, IL-10-producing CD4 + T cells, Tregs, Foxp3+ cells in brain, mLN, spleen (T7) MM: NA MC: EAE suppressed in recipient mice (mLN cells from T7) & depletion of CD4+ CD25 + T cells from mLN cells reverses suppression |
| Sanchez et al.49 | UN | M & F C57BL6 (WT, CD45.1, CD45.2, GF, TLR-2-/-, & TLR-9-/-) mice & SJL mice 9–10 wko n ≥ 10/g |
MOG-induced EAE (C57BL6) & PLP-induced EAE (SJL) |
T1: Proph L. paracasei ATCC 27092 in C57BL6 mice T2: Proph HK T1 in C57BL6 mice T3: Proph L. paracasei DSM 2649 in C57BL6 mice T4: Proph L. paracasei ATCC 11582 in C57BL6 mice T5: Proph L. paracasei ATCC 334 in C57BL6 mice T6: Proph L. paracasei DSM 5622 in C57BL6 mice T7: T2 in CD45.1 C57BL6 donor mice AT into CD45.2 C57BL6 mice T8: T2 in TLR-2-/- C57BL6 mice T9: T2 in TLR-9-/- C57BL6 mice T10: T2 in WT C57BL6 mice GMT into GF C57BL6 mice T11: Thera T2 in SJL mice Each 109 CFU o.g. C: PBS, MRS medium, or T1-conditioned medium |
Proph & Thera | Daily for 34–39 d (starting 2 wks pre-EAE) or daily for 39 d (starting 21 dpi) T10: daily for 28 d (starting 70 d pre-EAE) and GMT at 42 d pre-EAE |
CD: ↓ EAE CS (T1-T6); ↓ EAE incidence (T1-T6); ↓ demyelination, infiltrating macrophages & lymphocytes in brain & spinal cord, and severity of subsequent relapses (T1-T2); ↑ EAE CS w/ T1-conditioned media; – BBB & BSCB permeability (T2) IM: – prop. CD4+ IFN-γ + T cells, CD4+ IL-17A+ T cells, or Tregs in CNS (T1-T2); ↓ CCL3, CCL4, CXCL5, CXCL13 (T2) MM: ↓ EAE incidence (T10); – EAE CS (T10) MC: EAE suppression and ↓ demyelination eliminated in TLR2-/- mice; – EAE incidence or severity in AT recipient mice of T2 |
| Yamashita et al.67 | UN | F SJL mice 5 wko n = 10/g |
PLP-induced EAE |
T: HK L. helveticus SBT2171, 1 mg i.p. C1: 1 mg PBS i.p. C2: PBS, no EAE (n = 3) |
Proph & Thera | 3×/wk for 3 wks (pre-EAE), daily for 42 dpi |
CD: ↓ EAE incidence, CS, enlargement of inguinal LNs, infiltrating MNCs IM: ↓ # TH17, TH1, & CD4 + T cells in spinal cord, IL-17; ↓ IL-6, TGF-β, Foxp3+, IFN-γ in inguinal LNs; – IL-10 in inguinal LNs, – Ccl20 in spinal cord, CCR2, CCR4, & CCR6 on TH17 cells in dLNs MM: NA MC: NA |
| Libbey et al.54 | UN | M C57BL6 mice 4 wko n = 15/g |
MOG-induced EAE |
T1: rE. coli Nissle 1917, 1.31 × 109 CFU o.g. single gavage day T2: rE. coli Nissle 1917, 1.19 × 109 CFU + 1.25 × 109 CFU o.g. double gavage C1: 70 µL PBS C2: rE. coli Nissle 1917, no EAE |
Proph | 1–2 admins 3 or 7 d pre-EAE Follow for 35–36 dpi |
CD: ↓ survival & – CS, onset, incidence, CDS, maximal score, meningitis, demyelination, PVC (T1); ↓ CS, weight loss, PVC in T2; ↑ survival (T2); – EAE onset, incidence, CDS, maximal score, meningitis, demyelination (T2) IM: ↑ microglia, Tregs, IFN-γ, IL-27 (T1); ↓ CNS-derived cells, microglia, CD8 + T cells, CD4 + T cells, TH1 cells, Tregs in brain (T2); ↑ TH17 cells (T2) MM: NA MC: NA |
| Secher et al.55 | UN | M C57BL6 mice 8–12 wko n = 30–40/g |
MOG-induced EAE |
T1: E. coli Nissle 1917 T2: archetypal K12 E. coli MG1655 Each 108 CFU o.g. C: PBS |
Proph & Thera | Daily for 37 d (starting 7 d pre-EAE) |
CD: ↓ EAE CS, mortality, incidence, CDS, maximal score; – onset; ↓ colon & ileum permeability IM: ↓ total CD4+ and MOG-specific CD4 + T cells in spinal cord, IFN-γ, GM-CSF, IL-17, TNF-α, IL-6; ↑ total CD4+ and MOG-specific CD4 + T cells in LNs, IL-10, CD4+ Foxp3+ cells from draining LNs MM: NA MC: ↑ expression of Reg3g, Reg3b, Claudin-8, ZO-1 |
| Mestre et al.43 | UN | F SJL mice 5–8 wko n = 5–10/g |
TMEV-IDD |
T: TMEV-IDD w/ Vivomixxb 100 µL of 3 × 108 CFU o.g. C1: Sham w/ Vivomixxb C2: TMEV-IDD w/ vehicle C3: Sham w/ vehicle |
Thera | 3×/wk for 15 d (70–85 dpi) |
CD: ↑ horizontal & vertical activity, latency to fall IM: ↓ CD4 + T cells, B cells, % Foxp3+ CD39+ and Foxp3-CD39 + T cells in spleen, IL-1B, IL-6; ↑ Bregs, IL-10; – TNF-α; microglia exhibited anti-inflammatory activation or ↓ proinflammatory activity MM: ↓ rel. abund. of Anaerostipes, Dorea, Oscillospira, Enterobacteraceae, Ruminococcus, Bilophila, ↑ rel. abund. of Bacteroides, Odoribacter, Lactobacillus, Sutterella; ↑ acetate & butyrate in plasma MC: NA |
| Calvo-Barreiro et al.44 | Low | F C57BL6 OlaHsd mice 8 wko n = 17–20/g |
MOG-induced EAE |
T1: Lactibiane ikic, 1.6 × 109 CFU o.g. once daily T2: T1 twice daily T3: Vivomixxb, 9 × 109 CFU o.g. once daily T4: T3 twice daily C1/C2: water o.g. once or twice daily C3: untreated EAE C4: normal control |
Thera | 1–2× daily for 18–22 d (13–16 or 12–15 dpi) |
CD: ↓ CS w/ T1/T2 but not T3/T4 (dose-response observed); ↓ % demyelination, T cell inflammatory infiltrate density, & axonal damage (T1-T4); – intestinal permeability or microglia or astrocyte reactivity; improved motor coordination IM: ↓ ASP w/ T1/T2 but not T3/T4, % peripheral plasma cells (T1/T2); ↑ Tregs (T1/T2) MM: – alpha or beta diversity; ↑ rel. abund. of Lachnoclostridium and Bifidobacterium (T1/T2), Streptococcus (T3/T4); Atopobiacaeae & Bifidobacterium assoc. with ↓ accumulated EAE scores MC: 4x↓ expression of Th17 txn factor RORγt in spinal cord T1/T2 |
| Kwon et al.69 | UN | C57BL6 mice 6–8 wko n = 10/g |
MOG-induced EAE |
T1: Proph IRT5d T2: Thera IRT5d Each 5 × 108 CFU o.g. C: PBS |
Proph & Thera | Daily for 3 wks (pre-EAE) or 16 d (starting 12 dpi) |
CD: ↓ EAE incidence, CS, lymphocytes, Gr1+ and CD11b+ monocytes, and CD4 + T cells in spinal cord (T1); delayed EAE onset & ↓ EAE CS (T2) IM: ↓ ASP, IFN-γ, TNF-α, IL-17; ↑ IL-4, IL-10, CD4+ Foxp3+ Tregs from spinal cord MM: NA MC: NA |
| McMurran et al.45 | Low | F C57BL6 mice 13 mo n = 3–5/g |
Cuprizone (5 wks) |
T: VSL#3e, 1.35 × 109 CFU o.g. C: autoclaved water |
Proph & Thera | Daily for 7 wks (starting 4 wks pre-lysolecithin injection) |
CD: – remyelination IM: ↑ SCFAs in feces & serum, inflammatory response at 5 dpi, density of CD68+ activated microglia & infiltrating macrophages, # of ODCs at 5 dpi; – ODC response at 14 dpi, inflammatory response at 14 dpi MM: NA MC: NA |
| Ezendam et al.51 | UN | M & F Lewis rats 2 wko n = 4–8/g |
gpMBP-induced EAE |
T: B. animalis w/ EAE, 200 µL of 1 × 109 CFU o.g. C1: B. animalis w/ no EAE C2: 200 µL saline w/ EAE C3: 200 µL saline w/ no EAE |
Proph & Thera | Daily for ~60 d (starting 5 wks pre-EAE) |
CD: ↑ weight gain for M; shorter EAE duration for M; – EAE onset, duration, or CDI for F; – EAE onset or CDI for M IM: NA MM: NA MC: NA |
| Ezendam & van Loveren72 | High | M & F Lewis rats 2 wko n = 4–8/g |
gpMBP-induced EAE |
T; L. casei strain Shirota, 500 µL of 1–2 × 109 CFU o.g. C: 500 µL saline/peptone |
Proph & Thera | Daily for ~9 wks (starting 5 wks pre-EAE) |
CD: – EAE onset, CS, CDI for M & F; slightly longer duration for F; ↑ EAE incidence IM: NA MM: NA MC: NA |
| Johanson et al.76 | UN | F C57BL6 mice 8 wko n = 10/g |
MOG-induced EAE |
T: L. reuteri ATCC 2327 ad libitum C: MRS broth |
Thera | Daily for 20 d |
CD: ↓ average CS, – weight loss IM: NA MM: ↑ Lactobacillus 16S V3-V4 amplicon abundance MC: NA |
| Abdurasulova et al.66 | UN | F Wistar rats 3 mo n = 26–35/g |
homologous SCH-induced EAE |
T1: E. faecium LMG P-27496 L3 probe, 0.5 mL o.g. T2: Glatiramer acetate, 0.2 mL s.c. C1: saline, 0.5 mL o.g. C2: saline, 0.2 mL s.c. |
Thera | Daily for 15 d (starting 2 dpi) |
CD: ↓ prevalence, CDS, CS, mortality (T1/T2); delayed onset and shorter duration (T1/T2); T1 outperformed T2 in prevalence, duration, and CDS IM: ↓ CD4+ CD25+ Foxp3+ Tregs during peak and recovery phases, CD4 + T cells during peak; ↑ CD4+ Tcells during inductive, CD4+ CD25+ Foxp3- Tregs during peak, CD8 + T cells during peak & recovery MM: NA MC: NA |
All findings are reported with respect to control group(s) unless otherwise indicated.
bVivomixx = L. paracasei DSM 24734, L. plantarum DSM 24730, L. acidophilus DSM 24735, L. delbrueckii subsp. bulgaricus DSM 24734, B. longum DSM 24736, B. infantis DSM 24737, B. breve DSM 24732, S. thermophilus DSM 24731.
cLactibiane iki = B. lactis LA 304, L. acidophilus LA 201, L. salivarius LA 302.
dIRT5 = L. casei, L. acidophilus, L. reuteri, B. bifidum, S. thermophilus.
eVSL#3 = see bVivomixx.
Key: ↓ decreased; ↑ increased; – no change or no difference compared to control; NA, not applicable to this study.
Abbreviations: ROB, risk of bias; MS, multiple sclerosis; UN, uncertain; M, male; F, female; wk/wks/wko, week/weeks/weeks old; mo/mos, month/months; g, group; WT, wild-type; IL, interleukin; GF, germ-free; TLR, toll-like receptor; EAE, experimental autoimmune encephalomyelitis; MOG, myelin oligodendrocyte glycoprotein; PLP, proteolipid protein; MBP, myelin basic protein; gp, guinea pig; SCH, spinal cord homogenate; GID, gliotoxin-induced demyelination; TMEV-IDD, Theiler’s murine encephalomyelitis virus-induced demyelinating disease; CFU, colony-forming units; T, treatment group; C, control; o.g., oral gavage; IU, international units; w/w, weight/weight; HK, heat-killed; -/-, deficient; AT, adoptive transfer; GMT, gut microbiome transfer; i.p., intraperitoneally; PBS, phosphate-buffered saline; Proph, prophylactic; Thera, therapeutic; admins, administrations; dpi, days post immunization; CD, clinical disease; IM, immune/metabolic; MM, microbiome/metabolome; MC, mechanistic/correlative; CDB, clinical disease burden; CS, clinical score; CDI, clinical disease index; MNC, mononuclear cell; IgG, immunoglobulin G; ASP, antigen-specific proliferation; IFN-γ, interferon gamma; TGF-β, transforming growth factor beta; CDS, cumulative disease score; rel. abund., relative abundance; TNF-α, tumor necrosis factor alpha; CNS, central nervous system; LN, lymph node; BBB, blood–brain barrier; BSCB, blood spinal cord barrier; PVC, perivascular cuffing; GM-CSF, granulocyte monocyte colony-stimulating factor; txn, transcription; SCFA, short-chain fatty acid; ODC, oligodendrocyte; abs., absolute