Human Biofield Therapy and the Growth of Mouse Lung Carcinoma

Abstract

Biofield therapies have gained popularity and are being explored as possible treatments for cancer. In some cases, devices have been developed that mimic the electromagnetic fields that are emitted from people delivering biofield therapies. However, there is limited research examining if humans could potentially inhibit the proliferation of cancer cells and suppress tumor growth through modification of inflammation and the immune system. We found that human NSCLC A549 lung cancer cells exposed to Sean L. Harribance, a purported healer, showed reduced viability and downregulation of pAkt. We further observed that the experimental exposure slowed growth of mouse Lewis lung carcinoma evidenced by significantly smaller tumor volume in the experimental mice (274.3 ± 188.9 mm³) than that of control mice (740.5 ± 460.2 mm³; P < .05). Exposure to the experimental condition markedly reduced tumoral expression of pS6, a cytosolic marker of cell proliferation, by 45% compared with that of the control group. Results of reversed phase proteomic array suggested that the experimental exposure downregulated the PD-L1 expression in the tumor tissues. Similarly, the serum levels of cytokines, especially MCP-1, were significantly reduced in the experimental group ( P < .05). Furthermore, TILs profiling showed that CD8⁺/CD4^- immune cell population was increased by almost 2-fold in the experimental condition whereas the number of intratumoral CD25⁺/CD4⁺ (T-reg cells) and CD68⁺ macrophages were 84% and 33%, respectively, lower than that of the control group. Together, these findings suggest that exposure to purported biofields from a human is capable of suppressing tumor growth, which might be in part mediated through modification of the tumor microenvironment, immune function, and anti-inflammatory activity in our mouse lung tumor model.

Keywords: Lewis lung carcinoma; PD-L1; biofield; immune modulation.

Figure 1.

The effect of the experimental exposure (Ex) on the growth of human and mouse lung cancer cells. (A) The viability of human NSCLC A549 cells (Ex or control) measured 30 minutes and 3 hours after the exposures (3 hours). The Ex exposure led to a significant reduction of cell viability in this particular cell line. Cell viability was detected by PrestoBlue staining. (B) The viability of mouse Lewis Lung Carcinoma (LLC) cells 3 hours after the cells were exposed to Ex or control conditions measured by MTT assay. (C) The staining of mitotracker dye in A549 cells in control (a and b) and Ex (c and d) conditions. (D) Expression of pAkt and pERK in A549 cells at the end of the exposure (0.5 hour) or 3 hours after the Ex or control (Con) exposure. Data are presented as means ± standard error from 2 replicated experiments.

Figure 2.

The effect of the experimental exposure (Ex) on tumor growth in a mouse LLC model. (A) The tumor growth curves for Ex (n = 5) and control (n = 4) mice with LLC in which the exposures were initiated when tumors were barely palpable (experiment #1). (B) The tumor growth curves for Ex (n = 9) and control (n = 9) mice in which exposures were initiated when tumor volumes were 4 to 6 mm³ (experiment #2). The red arrows indicate when the mice received the experimental and control exposure. (C) The final tumor volumes in the Ex and control exposure mice from experiment #2. (D) The mean tumor weights in Ex and control exposure groups from the pooled data from experiments 1 and 2 (n = 13-14/group). (E) The tumor growth curves for Ex (n = 4) and control (n = 4) mice in which the exposures were initiated when tumor volume reached about 80 to 100 mm³ (experiment #3). The red arrow indicates when the mice received the experimental and control exposure. *P < .05, ***P < .005, for Ex exposure mice compared with controls. Data are presented as means ± standard error.

Figure 3.

Immunohistochemical staining of LLC sections for proliferation and apoptotic markers. Stains of tumor sections obtained from (A) control and (B) experimental exposure (Ex) mice for the cell proliferation marker Ki67. (C) Quantification of Ki67-positive cells in the tumor sections. Stains of tumor sections obtained from (D) control and (E) Ex mice for cleaved caspase-3. The red arrow indicates apoptotic cells. (F) Quantification of cleaved caspase-3-positive cells in the tumor sections. Stains of tumor section obtained from (G) control and (H) Ex mice for pS6 protein. (I) Quantification of pS6 positive cells in the tumor sections.

Figure 4.

PD-L1 expression in LLCs and serum chemokine and cytokine levels. (A) PD-L1 expression in tumor samples measured by RPPA. (B) Western blot of PD-L1 expression in the tumors from study mice. (C) Quantitative data for the Western blot results. (D) The serum levels of cytokines and chemokines in the control and Ex exposure mice. MCP-1, monocyte chemoattractant protein-1; mKC, mouse keratinocyte-derived chemokine; TNF-α, tumor necrosis factor α; IFN-g, interferon-γ.

Figure 5.

Immune modification in mice with LLC. (A) Tumors and (B) spleens of mice with LLC. The data suggest that experimental exposure (Ex) increased the number of CD8+ cytotoxic T cells while suppressing the number of regulatory T (Treg) cells, which are known to support the growth of tumor cells. Stains of tumor-infiltrating macrophages from (C) control and (D) Ex exposure mice for CD68. (E) Quantification of CD68+ cells in the tumors of the control and Ex exposure mice.