City of Hope Fibro Study
Dr. R. Paul St. Amand, Cladia Craig Marek, Dr John (Jack) Shively PhD and a team of researchers working on the "City of Hope Fibromyalgia Study" have e-published a paper ahead of print in the journal Experimental biology and medicine. 
Dr. R. Paul St. Amand is the originator of the Guaifenesin Protocol for Fibromyalgia Syndrome (Fibro), a much debated treatment protocol that is not widely accepted by the medical community. Claudia Craig Marek, his medical assistant, is the author of 'Fibromyalgia: The First Year'. The "City of Hope Fibromyalgia Study" is a three-year investigation involving patients of Dr St Amand, which is taking place at City of Hope Hospital in California, USA, where Dr Shively PhD is Chair of the Division of Immunology.
The purpose of the City of Hope Fibro study is to compare the blood of Fibro patients with that of healthy participitants in terms of autoinflammatory genes (genes involved with an inflammatory response when there is no obvious infection) and response to inflammatory stimuli. Other purposes are to determine if there are polymorphisms of the autoinflammatory genes and to look for evidence of links between Fibro and immune or genetic factors.
In the study discussed in the recently e-published article, blood plasma levels of 25 cytokines and chemokines in 92 female Fibro patients and 69 family members were measured, compared to 77 control samples taken from the City of Hope blood bank. The patient group included: patients whose parents did not have Fibro; patients who had one parent with Fibro; patients who had one parent with Fibro but a sibling that did not; and, in one case, a patient whose mother and maternal grandmother both had Fibro. Fifty-six (61%) of the patient group had been treated with the Guaifenesin Protocol for at least 3 months.  The control samples were known to be from females without Fibro, although it is not known how well they were screened for signs of the condition, or other conditions.
Cytokines are extensively within the body for communication between cells, like hormones and neurotransmitters, but with a greater diversity than either hormones (which are usually transported in the blood) or neurotransmitters (which are related to the nervous system). Cytokines are involved in the development and functioning of the immune system, as well as with a variety of immunological, inflammatory and infectious diseases. Chemokines are a group of small Cytokines, some of which are considered to promote inflammation.
The movement of leukocytes, a type of white blood cell, across membranes within the body, and the cytokine profile of skeletal muscle cells, were also analysed.
The study found that the Fibro patients and their family members had high levels of MCP-1 and eotaxin compared to controls.  However, it was not known how many of the family members might also have had Fibro themselves.
MCP-1, or Monocyte chemotactic protein-1, is a chemokine with a role in getting various immune system cells to the site of infection or injurys. Eotaxin is another chemokine, with a role in recruiting eosinophils, other immune system cells with a relation to inflammation, which are responsible for combating infection and parasites and also control mechanisms associated with allergy and asthma. MCP-1 and Eotaxin gene polymorphisms have been reported to contribute to susceptibility to several immune and inflammatory conditions.
The City of Hope study article also said that they had found that the patient group on the Guaifenesin Protocol had higher levels of eotaxin than those not being treated. 
The article says that "[d]iluted plasma from patients increased the migration of normal eosinophils and monocytes, but not neutrophils, through an endothelial/Matrigel barrier only when mast cells are included in the lower wells".  In other words, when compared to the controls, the samples from Fibro patients increased the amount of eosinophils and monocytes crossing barriers similar to those surrounding the internal vessels of the body, such as blood vessels: i.e. they increased the rate of movement of eosinophils and monocytes.
The study also found that skeletal muscle cells can, when tested in the laboratory (in vitro), secrete MCP-1, eotaxin, and IP-10, while treatment with MCP-1 caused secretion of IL-1beta, eotaxin and IP-10 (another cytokine). 
In the discussion, the researchers note that there are some possibly conflicting findings in their results, notably the raised levels of both MCP-1 and eotaxin. As eotaxin is a natural antagonist for the main receptor (CCR2) for MCP-1, the raised levels of eotaxin may be in response to the high levels of MCP-1 found, as the body tries to offset the damage caused by the MCP-1. This could also explain the high levels of IP-10 found, as IP-10 is the natural antagonist for both MCP-1 and eotaxin.
The researchers concluded that:
"the [Fibro group] studied here is associated with elevated levels of inflammatory chemokines, MCP-1 and eotaxin may contribute to the symptoms of [Fibro], parents of these patients share the expression profile, and [muscle cells] are a potential source of eotaxin and MCP-1. Therefore reduction of eotaxin and MCP-1 levels or blockade of their receptors may be a reasonable treatment strategy for [Fibro]. We also found evidence that treatment with guifenesin accentuates eotaxin production and reduces IL-13 and IFN-gamma [(other cytokines/chemokines)]. While only a few studies have shown elevated cytokine levels in [Fibro], our data suggests that elevated chemokine levels may play a causative role in [Fibro] and should be investigated further."
However, Hellhammer et al  found that chronic stress, such as that experienced by chronic pain patients, e.g. Fibro patients, disrupts the HPA axis and leads to low levels of cortisol. In the long-term, the low cortisol levels may lead to increased levels of other substances, such as leukocytes and cytokines.  Thus it could be argued that the raised levels of cytokines found in the Fibro patients in this study may be caused by the Fibro, rather than causing the Fibro in the first place. Or it could be a chicken-and-egg situation where the one worsens the other, making the first worse, and it is very hard to separate cause from effect.
There may also be a fault in the way the results are compared, which would mean that the raised levels of eotaxin in the patient group on the Guaifenesin Protocol compared to the patient group not on Guaifenesin may not be significant.
In summary, the study found that people with Fibro, whether they were on the Guaifenesin protocol or not, had high levels of substances that are usually associated with autoimmune and inflammatory conditions, suggesting that they may be an inflammatory or autoimmune connection with Fibro. Family members of the Fibro patients also had raised levels of some of these substances, suggesting that Fibro might have a family or genetic link. Dr St Amand has suggested that the raised levels of one of the immune system substances found in the patients on the Guaifenesin Protocol, as opposed to those who were not, may be of significance for the validity of the Guaifenesin Protocol. However, there were some flaws in the study and possibly in the conclusion, such as it not being known whether the family members themselves had Fibro and the difference between the Fibro patients on the Guaifenesin Protocol and those who were not, not in itself being significant.
This study remains a very interesting piece of research, but its significance may unfortunately be diluted by the flaws in the study and its significance to proponents of the Guaifenesin Protocol as evidence of the protocol is debatable.
- Zhang Z, Cherryholmes G, Mao A, Marek C, Longmate J, Kalos M, St Amand RP, Shively JE. High plasma levels of MCP-1 and eotaxin provide evidence for an immunological basis of fibromyalgia. Exp Biol Med (Maywood). 2008 Jun 5. [Epub ahead of print]
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- Chae SC, Park YR, Shim SC, Lee IK, Chung HT. Eotaxin-3 gene polymorphisms are associated with rheumatoid arthritis in a Korean population. Hum Immunol. 2005 Mar;66(3):314-20.
- Bugeja MJ, Booth D, Bennetts B, Heard R, Rubio J, Stewart G. An investigation of polymorphisms in the 17q11.2-12 CC chemokine gene cluster for association with multiple sclerosis in Australians. BMC Med Genet. 2006 Jul 26;7:64.
- Hellhammer DH, Ehlert U, Heim C. The potential role of hypocortisolism in the pathophysiology of stress-related bodily disorders. Psychoneuroendocrinology 25 (2000) 1–35