Bacterial lysate “Lantigen B” induces proliferation of B and NK cells and the release of related cytokines
DOI:
https://doi.org/10.65746/jbrha77Keywords:
immunostimulants; respiratory tract infections; Th1 immune reaction; cytotoxicity; trained immunityAbstract
Background: The present is an exploratory study, to evaluate the immune effect of a bacterial chemical lysate: Lantigen-B (Lan-B). PBMCs were stimulated with the concentration of the drug utilized in the clinical setting to evaluate its effects on lymphocyte subpopulation and cytokine release. Methods: PBMCs, from 7 healthy donors, were cultured alone or incubated with Lan-B at 199.45 Antigen Units/mL concentrations. Lymphocyte subpopulations were evaluated by Facs-Scan and cytokines by Cytometer. Results: A significant increase of IL-1b, IFN-g, TNF-a, IL-17, and IL-18 release and of CD19+ and CD16+/56+cells proportion was found in stimulated compared to unstimulated cultures along with a significant decrease of MCP1 and a trend for an increase of CD8+ cells and IL12-p40. Conclusions: The study shows that Lantigen-B drives a Th1-reaction and stimulates the differentiation/activation of B and NK cells. Changes in lymphocyte subpopulations and in cytokines are well correlated. The observed changes can result in a state of “prealert” of the immune system able to successfully fight infections also induced by bacteria and viruses different from those administered, the so-called “Trained Immunity”. Finally, the reduction of MCP1, which drives a Th2 reaction, can explain the beneficial effects on allergies observed in some clinical studies.
References
1. Suárez N, Ferrara F, Rial A, et al. Bacterial Lysates as immunotherapies for respiratory infections: Methods of preparation. Frontiers in Bioengineering and Biotechnology. 2020; 8: 545. doi: 10.3389/fbioe.2020.00545
2. Berber A, Del-Río-Navarro BE, Reyes-Noriega N, et al. Immunostimulants for preventing respiratory tract infection in children: A systematic review and meta-analysis. World Allergy Organization Journal. 2022; 15: 100684. doi: 10.1016/j.waojou.2022.100684
3. Huang Y, Pei Y, Qian Y, et al. A meta-analysis on the efficacy and safety of bacterial lysates in chronic obstructive pulmonary disease. Frontiers in Medicine. 2022; 9: 877124. doi: 10.3389/fmed.2022.877124
4. Braido F, Melioli G, Nicolini G, et al. Sublingually administered bacterial lysates: Rationale, mechanisms of action and clinical outcomes. Drugs Context. 2024; 13: 2024-1-5. doi: 10.7573/dic.2024-1-5
5. Di Gioacchino M, Santilli F, Pession A. Is there a role for immunostimulant bacterial lysates in the management of respiratory tract infection? Biomolecules. 2024; 14(10): 1249. doi: 10.3390/biom14101249
6. Drake CH, Smith JE. Letter: Salivary antibody response to oral vaccine. Lancet. 1975; 2: 614–615. doi: 10.1016/ S0140 6736(75)90215-9
7. Pozzi E, Serra C. Efficacy of Lantigen B in the prevention of bacterial respiratory infections. Monaldi Archives for Chest Disease= Archivio Monaldi per le Malattie del Torace. 2004; 61: 19–27.
8. XiaoQi S, YuHong L, ShuiYing Q, et al. Clinical curative effect of Lantigen B on bronchial asthma children with recurrent respiratory tract infection (Chinese). Modern Preventive Medicine 2013; 40: 1433–1434
9. Braido F, Melioli G, Nicolini G, et al. Prevention of recurrent respiratory tract infections: A literature review of the activity of the bacterial lysate Lantigen B. European Review for Medical & Pharmacological Sciences. 2023; 27. 7756–7767. doi: 10.26355/eurrev_202308_33430
10. Dirienzo W, Merli A, Ciprandi G, et al. Risposta anticorpale salivare e sierica ad una vaccinoterapia orale (Italian). Arch Med Intern. 1984; 36: 2–8.
11. Pizzimenti C, D’Agostino A, Pirrello P, et al. The SARS-CoV-2 cellular receptor ACE2 is expressed in oropharyngeal cells and is modulated in vitro by the bacterial lysate lantigen B. Archives of Clinical and Biomedical Research. 2023; 7: 13–18. doi: 10.26502/acbr.50170315
12. Chen Y, Lu D, Churov A, Fu R. Research Progress on NK Cell Receptors and Their Signaling Pathways. Mediators Inflamm. 2020; 24: 2020:6437057. doi: 10.1155/2020/6437057
13. Elemam NM, Ramakrishnan RK, Hundt JE, et al. Innate Lymphoid Cells and Natural Killer Cells in Bacterial Infections: Function, Dysregulation, and Therapeutic Targets. Frontiers in Cellular and Infection Microbiology. 2021; 11: 733564. doi: 10.3389/fcimb.2021.733564
14. Maity PC. B-Cell Receptor Signaling: Methods and Protocols. Springer Nature; 2025.
15. Oosting M, Brouwer M, Vrijmoeth HD, et al. Borrelia burgdorferi is strong inducer of IFN-γ production by human primary NK cells. Cytokine. 2022; 155: 155895. doi: 10.1016/j.cyto.2022.155895
16. Konjević GM, Vuletić AM, Mirjačić Martinović KM, et al. The role of cytokines in the regulation of NK cells in the tumor environment. Cytokine 2019; 117: 30–40. doi: 10.1016/j.cyto.2019.02.001
17. Schoenborn JR, Wilson CB. Regulation of interferon-gamma during innate and adaptive immune responses. Advances in Immunology. 2007; 96: 41–101. doi: 10.1016/S0065-2776(07)96002-2
18. Lei X, de Groot DC, Welters MJP, et al. CD4(+) T cells produce IFN-I to license cDC1s for induction of cytotoxic T-cell activity in human tumors. Cellular & Molecular Immunology. 2024; 21(4): 374–392. doi: 10.1038/s41423-024-01133-1
19. Lin Z, Zou S, Wen K. The crosstalk of CD8+ T cells and ferroptosis in cancer. Frontiers in Immunology. 2024; 14: 1255443. doi: 10.3389/fimmu.2023.1255443
20. Qiu Y, Tang M, Zeng W, et al. Clinical findings and predictive factors for positive anti-interferon-γ autoantibodies in patients suffering from a non-tuberculosis mycobacteria or Talaromyces marneffei infection: A multicenter prospective cohort study. Scientific Reports. 2022; 12(1): 9069. doi: 10.1038/s41598-022-13160-x
21. de Gruijter NM, Jebson B, Rosser EC. Cytokine production by human B cells: role in health and autoimmune disease. Clinical and Experimental Immunology. 2022; 210: 253–262, doi: 10.1093/cei/uxac090
22. Perez-Lopez A, Hernandez-Galicia G, Lopez-Bailon LU, et al. Pro-inflammatory and anti-inflammatory responses in B cells during Salmonella infection. European Journal of Microbiology and Immunology. 2025; 15(1): 32–41. doi: 10.1556/1886.2024.00088
23. Chang YP, Chen CL, Chen SO, et al. Autophagy facilitates an IFN-γ response and signal transduction. Microbes Infect. 2011; 13: 888–894. doi: 10.1016/j.micinf.2011.05.008
24. Afzal S, Abdul Manap AS, Attiq A, et al. From imbalance to impairment: the central role of reactive oxygen species in oxidative stress-induced disorders and therapeutic exploration. Frontiers in Pharmacology. 2023; 14: 1269581. doi: 10.3389/fphar.2023.1269581
25. Boehm U, Klamp T, Groot M, et al. Cellular responses to interferon-γ. Annual Review of Immunology. 1997; 15: 749–795. doi: 10.1146/annurev.immunol.15.1.749
26. Schroder K, Hertzog PJ, Ravasi T, et al. Interferon-γ: An overview of signals, mechanisms and functions. Journal of Leucocyte Biology. 2004; 75: 163–189. doi: 10.1189/jlb.0603252
27. Green AM, DiFazio R, Flynn JL. IFN-γ from CD4 T cells is essential for host survival and enhances CD8 T cell function during Mycobacterium tuberculosis infection. The Journal of Immunology. 2013; 190: 270–277. doi: 10.4049/jimmunol.1200061
28. Baird NL, Bowlin JL, Hotz TJ, et al. Interferon gamma prolongs survival of varicella-zoster virus-infected human neurons in vitro. Journal of Virology. 2015; 89: 7425–7427. doi: 10.1128/JVI.00594-15
29. Wu S, Zhang X, Hu C, et al. CD8(+) T cells reduce neuroretina inflammation in mouse by regulating autoreactive Th1 and Th17 cells through IFN-gamma. European Journal of Immunology. 2023; 53(12): e2350574. doi: 10.1002/eji.202350574
30. Borges da Silva H, Fonseca R, Alvarez JM, et al. IFN-γ Priming effects on the maintenance of effector memory CD4+T cells and on phagocyte function: Evidences from infectious diseases. Journal of Immunology Research, 2015; 2015(1): 202816. doi:10.1155/2015/202816
31. Falvo JV, Tsytsykova AV, Goldfeld AE. Transcriptional control of the TNF gene. Current Directions in Autoimmunity. 2010; 11: 27–60. doi: 10.1159/000289196
32. Idriss HT, Naismith JH. TNF alpha and the TNF receptor superfamily: Structure-function relationship(s). Microscopy research and technique. 2000; 50: 184–195.doi: 10.1002/1097-0029(20000801)50:3<184::aid-jemt2>3.0.co;2-h
33. Vallés G, Bensiamar F, Maestro-Paramio L, et al. Influence of inflammatory conditions provided by macrophages on osteogenic ability of mesenchymal stem cells. Stem Cell Research & Therapy. 2020; 11(1): 57. doi: 10.1186/s13287-020-1578-1
34. Alam MS, Otsuka S, Wong N, et al. TNF plays a crucial role in inflammation by signaling via T cell TNFR2. Proceedings of the National Academy of Sciences. 2021; 14: 118(50): e2109972118. doi: 10.1073/pnas.2109972118
35. Skartsis N, Ferreira LMR, Tang Q. The dichotomous outcomes of TNFα signaling in CD4+ T. Frontiers in Immunology. 2022; 13: 1042622. doi: 10.3389/fimmu.2022.1042622cells
36. Shapouri-Moghaddam A, Mohammadian S, Vazini H, et al. Macrophage plasticity, polarization, and function in health and disease. Journal of Cellular Physiology. 2018; 233: 6425–6440. doi: 10.1002/jcp.26429
37. Delves PJ, Roitt IM. The immune system: Second of two parts. New England Journal of Medicine. 2000; 343(1): 37–49. doi: 10.1056/NEJM200007133430207
38. Boyle JJ, Weissberg PL, Bennett MR. Tumor necrosis factor-α promotes macrophage-induced vascular smooth muscle cell apoptosis by direct and autocrine mechanisms. Arteriosclerosis, Thrombosis, and Vascular Biology. 2003; 23: 1553–1558. doi: 10.1161/01.atv.0000086961.44581.b7
39. Chen J, Jacobs-Helber SM, Barber DL, et al. Erythropoietin-dependent autocrine secretion of tumor necrosis factor-alpha in hematopoietic cells modulates proliferation via MAP kinase-ERK-1/2 and does not require tyrosine docking sites in the EPO receptor. Experimental Cell Research. 2004; 298: 155–166. doi: 10.1016/j.yexcr.2004.04.009
40. Lehner M, Kellert B, Proff J, et al. Autocrine TNF is critical for the survival of human dendritic cells by regulating BAK, BCL-2, and FLIP L. The Journal of Immunology. 2012;188: 4810–4818. doi: 10.4049/jimmunol.1101610
41. Innins EK, Gatanaga M, Eden M, et al. The autocrine role of tumor necrosis factor in the proliferation and functional differentiation of human lymphokine-activated T killer cells (T-LAK) in vitro. Cytokine. 1992; 4: 391–396. doi: 10.1016/1043-4666(92) 90083- 4
42. Rychly DJ, DiPiro JT. Infections Associated with Tumor Necrosis Factor-α Antagonists. Pharmacotherapy. 2005; 25(9): 1181–1192.doi: 10.1592/phco.2005.25.9
43. You K, Gu H, Yuan Z, et al. Tumor necrosis factor alpha signaling and organogenesis. Frontiers in Cell and Developmental Biology. 2021; 30: 9:727075. doi: 10.3389/fcell.2021.727075
44. Boussiotis VA, Nadler LM, Strominger JL, et al. Tumor necrosis factor-a is an autocrine growth factor for normal human B cells. Proceedings of the National Academy of Sciences. 1994; 91: 7007–7011 doi: 10.1073/pnas.91.15.7007
45. Bouwmeester T, Bauch A, Ruffner H, et al. A physical and functional map of the human TNF-α/NF-kappa B signal transduction pathway. Nature Cell Biology. 2004; 6: 97–105. doi: 10.1038/ncb1086
46. Jelinek DF, Lipsky PE. Enhancement of human B cell proliferation and differentiation by tumor necrosis factor-alpha and interleukin 1. The Journal of Immunology. 1997; 139: 2970–2976. doi: 10.4049/jimmunol.139.9.2970
47. Wewers MD, Dare HA, Winnard AV, et al. IL-1 beta-converting enzyme (ICE) is present and functional in human alveolar macrophages: macrophage IL-1 beta release limitation is ICE independent. The Journal of Immunology. 1997; 159: 5964–5972. doi: 10.4049/jimmunol.159.12.5964
48. Yaseen MM, Abuharfeil NM, Darmani H. “The role of IL-1β during human immunodeficiency virus type 1 infection”. Reviews in Medical Virology. 2023; 33(1): e2400. doi: 10.1002/rmv.2400
49. Lopez-Castejon G, Brough D. Understanding the mechanism of IL-1beta secretion. Cytokine & Growth Factor Reviews. 2011; 22: 189–195. doi: 10.1016/j.cytogfr.2011.10.001
50. Oberholzer A, Oberholzer C, Moldawer LL. Cytokine signaling—regulation of the immune response in normal and critically ill states. Critical Care Medicine. 2000; 28(4 Suppl.): N3–N12. doi: 10.1097/00003246-200004001-00002
51. Andrei C, Dazzi C, Lotti L, et al. The secretory route of the leaderless protein interleukin 1β involves exocytosis of endolysosome- related vesicles. Molecular Biology of the Cell. 1999; 10: 1463–1475. doi: 10.1091/mbc.10.5.1463
52. Sitia R, Rubartelli A. The unconventional secretion of IL-1β: Handling a dangerous weapon to optimize inflammatory responses. Seminars in Cell & Developmental Biology. 2018; 83: 12–21. doi: 10.1016/j.semcdb.2018.03.011
53. Semino C, Carta S, Gattorno M, et al. Progressive waves of IL-1β release by primary human monocytes via sequential activation of vesicular and gasdermin D-mediated secretory pathways. Cell Death & Disease. 2018; 9:1088. doi: 10.1038/s41419-018-1121-9
54. Nakae S, Masahide Asano M, Horai R, et al. Interleukin-1β, but not interleukin-1α, is required for T-cell-dependent antibody production. Immunol. 2001; 104: 402–409. doi: 10.1046/j.1365-2567.2001.01337.x
55. Van Den Eeckhout B, Tavernier J, Gerlo S. Interleukin-1 as Innate mediator of T cell immunity. Frontiers in Immunology. 2021; 11: 2020. doi: 10.3389/fimmu.2020.621931
56. Ihim SA, Abubakar SD, Zian Z, et al. Interleukin-18 cytokine in immunity, inflammation, and autoimmunity: Biological role in induction, regulation, and treatment. Frontiers in Immunology. 2022; 13: 919973. doi: 10.3389/fimmu.2022.919973
57. Mantovani A, Dinarello CA, Molgora M, et al. Interleukin-1 and Related Cytokines in the Regulation of Inflammation and Immunity. Immunity. 2019; 50: 778–795. doi: 10.1016/j.immuni.2019.03.012
58. Nakanishi K. Unique action of interleukin-18 on t cells and other immune cells. Frontiers in Immunology. 2018; 9: 763. doi: 10.3389/fimmu.2018.00763
59. Kaplanski G. Interleukin-18: Biological properties and role in disease pathogenesis. Immunol Review. 2018; 281: 138–153. doi: 10.1111/imr.12616
60. Nakanishi K, Yoshimoto T, Tsutsui H, et al. Interleukin-18 regulates both Th1 and Th2 responses. Annual Review of Immunology. 2001; 19: 423–474. doi: 10.1146/annurev.immunol.19.1.423
61. Tsutsui H, Nakanishi K, Matsui K, et al. IFN-gamma-inducing factor up-regulates Fas ligand-mediated cytotoxic activity of murine natural killer cell clones. The Journal of Immunology. 1996; 157: 3967–3973. doi: 10.4049/jimmunol.157.9.3967
62. Madera S, Sun JC. Cutting edge: stage-specific requirement of IL-18 for antiviral NK cell expansion. The Journal of Immunology. 2015; 194: 1408–1412. doi: 10.4049/jimmunol.1402001
63. Freeman BE, Raue HP, Hill AB, et al. Cytokine-mediated activation of NK cells during viral infection. Journal of Virology. 2015; 89: 7922–7931. doi: 10.1128/ JVI.00199-15
64. Abdi K. IL-12: The role of p40 versus p75. Scandinavian Journal of Immunology. 2002; 56: 1–11. doi: 10.1046/j.1365-3083.2002.01101.x
65. Tait Wojno ED, Hunter CA, Stumhofer JS. The Immunobiology of the Interleukin-12 Family: Room for Discovery. Immunity. 2019; 50: 851–870. doi: 10.1016/j.immuni.2019.03.011
66. Abel AM, Yang C, Thakar MS, et al. Natural killer cells: Development, maturation, and clinical utilization. Frontiers in Immunology. 2018; 9: 1869. doi: 10.3389/fimmu.2018.01869
67. Landoni, E. et al. IL-12 reprograms CAR-expressing natural killer T cells to long-lived Th1-polarized cells with potent antitumor activity. Nature Communications. 2024; 15: 89. doi: 10.1038/s41467-023-44310-y
68. Clark JT, Weizman OE, Aldridge DL, et al. IL-18BP mediates the balance between protective and pathological immune responses to Toxoplasma gondii. Cell Reports. 2023; 42(3): 112147. doi: 10.1016/j.celrep.2023.112147
69. Wang H, Ruan G, Li Y, et al. The Role and Potential Application of IL-12 in the Immune Regulation of Tuberculosis. International Journal of Molecular Sciences. 2025; 26(7): 3106. doi: 10.3390/ijms26073106
70. Cheng Y, Luo R, Li E. Combined delivery of IL12 and an IL18 mutant without IL18BP-binding activity by an adenoviral vector enhances tumor specific immunity. Scientific Reports. 2025; 15(1): 3563. doi: 10.1038/s41598-024-84693-6
71. Wang Y, Chaudhri G, Jackson RJ, et al. IL-12p40 and IL-18 play pivotal roles in orchestrating the cell-mediated immune response to a poxvirus infection. The Journal of Immunology. 2009; 183(5): 3324-3331. doi: 10.4049/jimmunol.0803985
72. Pol JG, Workenhe ST, Konda P, et al. Cytokines in oncolytic virotherapy. Cytokine & Growth Factor Reviews. 2020; 56: 4–27. doi: 10.1016/j.cytogfr.2020.10.007
73. Huangfu L, Li R, Huang Y, et al. The IL-17 family in diseases: From bench to bedside. Signal Transduction and Targeted Therapy. 2023; 8(1): 402. doi: 10.1038/s41392-023-01620-3
74. Mills KHG. IL-17 and IL-17-producing cells in protection versus pathology. Nature Reviews Immunology. 2023; 23: 38–54. doi: 10.1038/s41577-022-00746-9
75. Dadaglio G, et al. IL-17 suppresses the therapeutic activity of cancer vaccines through the inhibition of CD8(+) T-cell responses. Oncoimmunology. 2020; 9: 1758606. doi: 10.1080/2162402X.2020.1758606
76. Al Omar S, Flanagan BF, Almehmadi M, et al. The effects of IL-17 upon human natural killer cells. Cytokine. 2013; 62: 123–130. doi: 10.1016/j.cyto.2013.02
77. Happel KI, Dubin PJ, Zheng M, et al. Divergent roles of IL-23 and IL-12 in host defense against Klebsiella pneumoniae. The Journal of Experimental Medicine. 2005; 202: 761–769. doi: 10.1084/jem.20050193c
78. Troy NM, Strickland D, Serralha M, et al. Protection against severe infant lower respiratory tract infections by immune training: mechanistic studies. Journal of Allergy and Clinical Immunology. 2022; 150: 93–103. doi: 10.1016/j.jaci.2022.01.001
79. Ricci R, Palmero C, Bazurro G, et al. The administration of a polyvalent mechanical bacterial lysate in elderly patients with COPD results in serological signs of an efficient immune response associated with a reduced number of acute episodes. Pulmonary Pharmacology & Therapeutics. 2014; 27: 109–113. doi: 10.1016/j.pupt.2013.05.006
80. Domínguez-Andrés J, Dos Santos JC, Bekkering S, et al. Trained immunity: adaptation within innate immune mechanisms. Physiological Reviews. 2023; 103(1): 313–346. doi: 10.1152/physrev.00031.2021
81. Ochando J, Mulder WJM, Madsen JC, et al. Trained immunity — basic concepts and contributions to immunopathology. Nature Reviews Nephrology. 2023; 19: 23–37. doi: 10.1038/s41581-022-00633-5
82. Acevedo OA, Berrios RV, Rodríguez-Guilarte L, et al. Molecular and cellular mechanisms modulating trained immunity by various cell types in response to pathogen encounter. Frontiers in Immunology. 2021; 4: 12: 745332. doi: 10.3389/fimmu.2021.745332
83. Teufel LU, Arts RJW, Netea MG, et al. IL-1 family cytokines as drivers and inhibitors of trained immunity. Cytokine. 2022; 150: 155773. doi: 10.1016/j.cyto.2021.155773
84. Martín-Cruz L, Benito-Villalvilla C, Angelina A, et al. Trained immunity-based vaccines for infections and allergic diseases. Journal of Allergy and Clinical Immunology. 2024; 154(5): 1085–1094. doi: 10.1016/j.jaci.2024.09.009
85. Bindu S, Dandapat S, Manikandan R, et al. Prophylactic and therapeutic insights into trained immunity: A renewed concept of innate immune memory. Human Vaccines & Immunotherapeutics. 2022; 18: 2040238. doi: 10.1080/21645515.2022.2040238
86. Singh S, Anshita D, Ravichandiran V MCP-1: Function, regulation, and involvement in disease. International Immunopharmacology. 2021; 101(Pt B): 107598. doi: 10.1016/j.intimp.2021.107598
87. Rihar M, Bahri R, Forstnerič V, et al. CCL2/C-C chemokine receptor type 2-mediated interactions among mast cells, basophils, and endothelial cells. Clinical and Translational Allergy. 2025; 15(2): e70044. doi: 10.1002/clt2.70044
88. Vercelli D. From Amish farm dust to bacterial lysates: The long and winding road to protection from allergic disease. Seminars in Immunology. 2023; 68: 101779. doi: 10.1016/j.smim.2023.101779
89. Kaczynska A, Klosinska M, Janeczek K, et al. Promising immunomodulatory effects of bacterial lysates in allergic diseases. Frontiers in Immunology. 2022; 13: 907149. doi: 10.3389/fimmu.2022.907149
90. Janeczek K, Emeryk A, Rachel M, et al. Polyvalent mechanical bacterial lysate administration improves the clinical course of grass pollen-induced allergic rhinitis in children: A randomized controlled trial. The Journal of Allergy and Clinical Immunology: In Practice. 2021; 9: 453–462. doi: 10.1016/j.jaip.2020.08.025
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