Author: Alan R Cantwell, Jr (---.vnnyca.adelphia.net)
Date: 08-26-03 23:03
Cell-Wall-Deficient Bacteria as a Possible Cause of Basal Cell Carcinoma
Author: Alan R. Cantwell, Jr., M.D., Los Angeles, CA
Received: 8 Aug 2003
Published: 26 August 2003
ABSTRACT
Basal cell carcinoma is the most common form of skin cancer, as well as the most common malignant tumor of human beings. The purpose of this study was to determine if so called "cancer microbes" could be demonstrated within the histopathologic tissue sections from this tumor. Variably acid-fast extracellular and intracellular coccoid forms, suggestive of cell-wall-deficient bacteria, were observed within the fibrous stroma and within the tumor cell nests in microscopic tissue sections from 35/37 cases (96%) of basal cell carcinomas studied. These coccoid and granular forms in vivo were similar in size and shape to coccal and granular elements of Staphylococcus epidermidis and Propionobacterium acnes isolated in thioglycollate broth culture from biopsy material of several basal cell carcinomas. The histologic presence of these coccoid and granular forms suggests that these microbes might be implicated in the pathogenesis of basal cell carcinoma. The idea of bacteria as causative agents in cancer is generally considered anathema by cancer researchers. However, continuing microbiologic studies suggest that latent hematologic infection with cell-wall-deficient bacteria may be present in the blood of all human beings. Just as viruses have been long thought to be implicated in the pathogenesis of cancer, it is not unreasonable to suspect that newly-discovered cryptic cell-wall-deficient bacteria harbored in the blood, along with their possible effect on the immune system, may play some pathogenic role in the production of cancer as well.
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Tiny purple-stained coccoid forms inside a cell of basal cell carcinoma.
Fite Stain, x1000, in oil (click here to enlarge image).
INTRODUCTION
The precise cause of basal cell carcinoma (BC ca) of the skin is not known. Chronic sun exposure, especially in fair-skinned persons, is a well-accepted factor in the development of skin cancer. However, BC ca may occasionally develop in patients on areas of skin that have never been exposed to the sun, such as the axilla, buttocks and vulva.
Over the past half-century, a number of different investigators have suggested that "cancer bacteria" in the form of variably acid-fast cell-wall-deficient (CWD) bacteria may be associated with cancer [1-4]. In this regard, Cantwell et al. have demonstrated variably acid-fast coccoid forms (suggestive of CWD bacteria) in microscopic sections of breast cancer, classic and AIDS-related forms of Kaposi's sarcoma, Hodgkin's disease, mycosis fungoides (T-cell lymphoma), as well as in immunologic diseases like lupus erythematosus and sarcoidosis, and in certain connective tissue diseases, such as scleroderma and pseudoscleroderma (hypodermitis sclerodermiformis) [5-13]. Cantwell's research is summarized in Cell Wall Deficient Bacteria (1982) and The Cancer Microbe (1990) [14, 15].
Bacteriologic studies by Pohlod et al.[16], Tedeschi et al.[17,18], and Domingue et al. [19], all indicate that human blood may be cryptically infected with CWD bacteria . In the late 1970s Tedeschi and other Italian microbiologists discovered microbial "granules" in the red blood cells of healthy and ill people. Some of the bacteria cultured from the blood cells were acid-fast [18], a feature shared with the cancer microbe. Cultures of these pleomorphic blood bacteria were identified as staphylococcal, streptococcal, corynebacteria and propionibacteria-like (diphtheroid) microbes.
Recent confirmatory reports by Brown et al. and McLaughlin et al. suggest that latent hematologic infection with CWD bacteria may be universal [20,21]. Findings of heretofore unsuspected blood bacterial pathogens lend new support to the century-old heretical theory that bacteria cause cancer. Furthermore, pleomorphic staphylococcal, corynebacteria-like, and propionibacteria-like microbes have been repeatedly cultured from tumors by cancer microbe researchers, and these isolates are similar to the kinds of pleomorphic bacteria now being isolated and identified from human blood.
The association of CWD microbes with BC ca is explored in this report.
MATERIALS AND METHODS
Acid-fast stained microscopic sections of 51 histologically proven tumors of BC ca were studied under oil-immersion magnification (x1000) for the presence of variably acid-fast coccoid and rod forms. Acid-fast staining procedures included the Ziehl-Neelsen, Fite, and the Intensified Kinyoun stain, recommended by Mattman for the demonstration of acid-fast CWD forms of mycobacteria in vivo [22]. A number of different investigators have previously noted that acid-fast staining procedures (rather than Giemsa or Gram staining) provide the most satisfactory differential staining method for the demonstration of cancer-associated bacteria in vivo.
The patient group consisted of 37 cases (25 males; 12 females), ranging in age from 25 to 81 years of age. All patients were caucasian. All lesions were located on sun-exposed areas (head, trunk, arms, and legs).
The bacteriology of BC ca was not determined in this study. However, preliminary results of bacteriologic culture (in thioglycollate broth) of several of these tumors, as well as "normal" surrounding skin, have yielded growth of pleomorphic organisms identified as Propionibacterium acnes, or as Staphylococcus epidermidis with unusual staining characteristics. Microphotographs of two bacterial isolates are presented in order for the reader to compare the morphologic of the cocci and granules isolated in vitro with these coccoid forms observed in vivo (Figs. 5-7). "Cancer microbes" isolated from malignant tumors have ordinarily been considered as "contaminants", or as "secondary invaders." For this reason, the emphasis in this report is on the demonstration of in vivo microbial forms. Unfortunately, there have been no modern bacteriologic investigations of tumor tissue in BC ca.
Microphotographs of six different histologic cases of BC ca were selected for presentation. Details of these cases are provided in Table 1.
TABLE 1
Case Sex Age Location Diagnosis
1 F 68 Right Temple BC ca (multiple)
2 M 61 Right scapular area BC ca
3 M 62 Left forehead BC ca
4 M 67 Glabella BC ca
5 M 59 Sternal area Recurrent BC ca
6 F 45 Left temple BC ca (multiple) malignant melanoma
COLOR MICROPHOTOGRAPHS
 
(click here to enlarge image 1A) (click here to enlarge image 1B)
1. Basal cell carcinoma. Fite stain, x1000, in oil.
(A) Extracellular purple-stained coccoid forms in center, with nearby
collection of intracellular coccod forms. Large pink bodies on right are red blood cells.
(B) Multiple foci of intra- and extracellular coccoid forms between nests of tumor cells.
 
(click here to enlarge image 1C) (click here to enlarge image 1D)
1. Basal cell carcinoma. Fite stain, x1000, in oil.
(C) Large focus of closely-packed coccoid forms left of center. Red blood cells at upper right.
(D) Multiple foci of densely-packed coccoid forms.
 
(click here to enlarge image 2A) (click here to enlarge image 2B)
2. Recurrent Basal cell carcinoma. Fite stain, x1000, in oil.
(A) Solitary focus of extracellular coccoid forms in center between collections of basal cell tumor cells.
(B) Large cell packed with coccoid forms. A few scattered coccoid forms are seen at the upper right.
RESULTS (photo references refer to b/w photos below)
Extracellular, variably acid-fast coccoid forms were noted in microscopic sections of BC ca derived from 35/37 patients (96%), and in 49/51 slides (96%). The two negative results were found in a BC ca lesion of the ala nasi occurring in a 63 year-old female; and in a posterior trunk lesion occurring in a 65 year-old male. Sections from three previously removed BC ca from the latter patient revealed extracellular coccoid forms with acid-fast staining.
Extracellular coccoid forms were defined as scattered forms or as closely-knit foci of coccoid forms clearly unassociated with a cell nucleus (Figures 1,4,5,6). These extracellular forms were most numerous within the fibrous stroma surrounding the BC ca tumor nests (Figure 2). Large numbers of coccoid forms appeared to be located intracellularly or in close proximity to a cell nucleus. Fewer coccoid forms were seen at the edges of the tumor where the epidermis became more normal histologically, and in the deep dermis underlying the tumor. Rare coccoid forms could also be detected in the dermis in biopsy sections of "normal" skin in close proximity to the BC ca (Figure 6).
DISCUSSION
The theory of an infectious ''cancer parasite" associated with cancer was totally discredited in the early years of the twentieth century. To this day, the idea of a "cancer microbe" is considered heresy by the cancer establishment; and any health professional who dares revive the idea is likely to be viewed as a quack.
For a century, there have been two main arguments against bacteria found in cancer. First, so-called "cancer microbes" are considered to be "laboratory contaminants" or "secondary bacterial invaders" of diseased tissue.
In this study, the "in vivo" demonstration of microbe-like elements within the tissue sections of BC ca indicates that coccoid and granular forms (irrespective of their "true" nature) are bona fide elements within the cancerous tissue and cannot be considered "contaminants" in any way. Second, these microbes appear in areas away from the tumor growth in areas of skin that are "normal" and cancer-free. This indicates that these microbe-like forms are present before the normal tissue becomes cancerous. By preceeding the development of cancer, they could be considered as "primary", but certainly not "secondary invaders."
Obviously, laboratory contaminants and secondary invaders must be ruled-out when bacteria are cultured from cancer, particularly when no bacteria can be observed in the tissue from which the bacteria were cultured. However, to automatically consider all bacteria (particularly staphylococci and corynebacteria) as contaminants of tumor tissue is no longer valid, simply because such bacteria are now considered to be bona fide infective elements within the blood — and because elements of microbe-like structures (coccoid forms and granules) can be identified in tumors, as shown in this report.
It would seem that all diseases in which the immune system plays a significant role would need to be re-evaluated now that blood is believed to carry occult and potentially pathogenic CWD bacteria. The comforting concept of the "sterility" of "normal and healthy" blood no longer seems scientifically valid. Thus, when microbes are cultured from cancer and other disease states — and when similar microbial elements are observed within the diseased tissue — they must be taken seriously as possible disease agents and investigated carefully.
The demonstration of intra- and extracellular variably acid-fast coccoid and granular forms in vivo in BC ca will undoubtedly spark controversy as to their precise nature. In 1948, microscopist Roy Allen wrote that "maximum proliferation of the organisms (associated with cancer in vivo) apparently occurs within the confines of a tumor cell. Sooner of later, however, the organisms break out from the confines of a cell and migrate to surrounding tissues. Multiplication may take place outside the cells, especially when collagenous tissue is present. It is probable that these organisms have been seen and noted by practically every pathologist but interpreted as eosinophilic granules (in hematoxylin-eosin stained sections), hence their true nature not recognized. Only the Ziehl-Neelsen (acid-fast stain) technique reveals the difference."
In consulting with pathologists over several decades, I was aware that they routinely interpreted these tiny acid-fast stained bodies as inconsequential mast cell granules of no great importance, even though mast cells are not known to be acid-fast. They simply could not accept the possibility that these tiny bodies were variably acid-fast CWD bacteria. As a result, no pathologist was ever willing to note these acid-fast forms in any of my patients' official biopsy reports. These doctors all knew I was investigating their tissue sections for the presence of bacteria in cancer and other diseases "of unknown etiology". Undoubtedly, they were reluctant to note any hint of a suspected infective bacterial element, particularly in cancer where the idea of cancer parasites is taboo. Clearly, no respectable pathologist wanted to break that taboo.
Thus, attempting to open the minds of histopathologists to the possibility that CWD bacteria might be hiding in tissue under the guise of mast cell granules is a Herculean task. Surely, a careful reading of the heretical microbiology of cancer, as proposed by Livingston, Alexander-Jackson, Seibert, Diller, and others, would go a long way in recognizing how "the cancer microbe" might appear in tissue, in blood, and in laboratory culture [1-4].
In their seminal 1950 paper describing the microscopic and electron microscopic appearance of microbes derived from the blood of cancerous patients, Wuerthele-Caspe Livingston and her associates describe how the tiny viral-like "granules" observed in culture increase in size as the culture ages, growing into larger round forms ("globoids") "until they become as large as staphylococci." [1] The researchers also called attention to the similarity of these forms in culture to the appearance of mast cells in tissue. The authors wrote: "The organisms appear to be embedded in irregular strands of collagen-like material. The minute acid-fast forms are present as well as aggregates of larger globoid bodies in this matrix. These aggregates strongly resemble groups of mast cells which have disintegrated and released their granules. It seems possible that certain granules which have not infrequently been observed within diseased tissues and which frequently have been called mast cell granules, may be globoidal forms of a microorganism of this type."
To further complicate the matter of distinguishing microbial granules from mast cell granules in fixed tissue sections is the difficulty in precisely defining the morphologic and tinctorial characteristics of a mast cell. In this regard, Simpson, in 1963, noted: "In the spirit of the confession that is alleged to be good for the soul, I admit that I have tried, but have never been able to define the term mast cell." Two decades later, the heteromorphic and heterogenic qualities of mast cells prompted an editorial by Kaliner, perhaps aptly-titled "Is a mast cell a mast cell a mast cell?". Suffice to say, the morphologic configurations presented as microbial forms in this paper do not resemble published microphotographs of dermal and intraepidermal mast cells as illustrated by James and Eady (their Figure 3) or by Leder [26,27].
Until the precise nature of the myriad of coccoid and granular forms both within the fibrous connective tissue and within the tumor cell nests in BC ca is ascertained, the precise relationship (if any) between these forms and mast cells granules must await further clarification and observations.
The finding of bacteria in the most common form of skin cancer confirms the contention of other researchers that a cancer microbe exists in cancer. Microphotographic evidence strongly suggests that the coccoid forms in BC ca bear a close resemblance to coccal elements of Propionibacterium acnes and Staphylococcus epidermidis isolated from these tumors, as shown in Figures 5-7. Admittedly, such bacteria can be cultured from "normal" skin, and are frequent laboratory contaminants as well. However, the existence of similar-sized forms within the tumor must allow some consideration of these microbes as potential pathogens.
The persistent claim by Wuerthele-Caspe Livingston et al. that the "cancer microbe" is distinguished in vivo and in vitro by its "intermittent acid-fastness" is in accord with the finding that the S. epidermidis isolated from a lesion of BC ca (Figure 7) was indeed partially acid-fast. Notably, the "typical" cocci in culture were not acid-fast, but the minute acid-fast "granules" and tiny rods were acid-fast. Ordinarily, cultures of staphylococci isolated from clinical material are not stained and observed for acid-fast elements.
BC ca is the most common and accessible malignant tumor of human beings. The further study of variably acid-fast coccoid elements in vivo, along with more detailed microbiologic and histopathologic investigations of BC ca, could increase our knowledge of the possible role of CWD bacteria in the pathogenesis of skin cancer and other forms of cancer.
LEGEND FOR MICROPHOTOGRAPHS
(click here to enlarge image).
Figure 1. (A) Microscopic section of basal cell carcinoma (BC ca) showing purple-stained extracellular coccoid forms near a tumor cell nest (arrows).
(B) Similar extracellular coccoid forms in the fibrous stroma along with a possible mast cell (mc) containing similar, tightly-packed coccoid forms (Case 1; Fite stain, magnification x1000, in oil).
(click here to enlarge image).
Figure 2. Microscopic section of BC ca showing intracellular coccoid forms within the tumor nest (Case 2; Ziehl-Neelsen stain, x1000, in oil).
(click here to enlarge image).
Figure 3. Section of BC ca showing extra- and intracellular coccoid forms (arrows) within the fibrous stroma of the upper dermis surrounding two basal cell (bc) nests (Case 3; Fite stain, x1000, in oil).
(click here to enlarge image).
Figure 4: Section of BC ca showing acid-fast coccoid forms (arrows) in the fibrous stroma (S) surrounding a basal cell (bc) nest (Case 4; Fite stain, x1000, in oil).
(click here to enlarge image).
Figure 5: (A) Recurrent BC ca showing numerous acid-fast coccoid forms (arrows) in the deep dermis (Case 5; Ziehl-Neelsen stain, x 1000, in oil).
(B) Additional section stained with the Brown-Brenn (Gram) stain showing very rare Gram-negative (pink-staining) coccoid forms along with tumor cells in the deep dermis.
(C) Smear of non-acid-fast P. acnes isolated from the same lesion. Note the size of the coccal elements and compare with the coccoid forms observed in vivo in (A) and (B) (Ziehl-Neelsen stain, x1000, in oil).
(click here to enlarge image).
Figure 6. (A) Section of BC ca showing extracellullar coccoid froms in the upper dermis in the stroma between two tumor cell nests (Case 6; Fite stain, x1000, in oil).
(B) Gram-stained smear from Gram-positive S epidermidis isolated from this lesion. Note the variability of the size of the cocci within the same culture.
(C) Minute coccoid forms observed in the deep dermis of "normal" skin near a BC ca in the same patient (Fite stain, x1000, in oil).
(click here to enlarge image).
Figure 7. Smear from culture of BC ca and identified as S epidermidis. Pleomorphic elements include non-acid-fast regular sized cocci (C), smaller non-acid-fast "micrococci" (mC), numerous tiny, acid-fast granules (g), and short acid-fast rods (r) (Case 6; Ziehl-Neelsen stain, x1000, in oil).
REFERENCES
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ACKNOWLEDGEMENTS
Dan W. Kelso, Chief, Bacteriology section. Central Diagnostic Laboratories, Tarzana, CA, performed the bacteriologic isolations reported in this study. This study was performed in 1982 at the Department of Dermatology, Southern California Permanente Medical Group, 1505 North Vermont Avenue, Los Angeles, CA 90027. The study was supported and approved by a research grant from the Medical Group. Dr. Cantwell is now retired from the practice of dermatology.
KEY WORDS
carcinoma, basal cell
bacteria, acid-fast
cell-wall-deficient bacteria
histopathologic findings
Staphylococcus epidermidis
Propionibacterium acnes
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